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2024-03-29T09:28:07Z
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http://2014.igem.org/Team:ITB_Indonesia/Parts
Team:ITB Indonesia/Parts
2014-10-18T02:11:52Z
<p>Erawijantari: </p>
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<div id="center"><br />
<div id="logo"></div><br />
<br />
<div id="header"></div><br />
<br />
<div id="blockNav"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>PARTS</h1><br />
<br><br />
<img src="https://static.igem.org/mediawiki/2014/d/db/Parts1.JPG"><br />
<br><br />
<br><br />
<h1>PARTS DESCRIPTION</h1><br />
<br><br />
<ol type="A"><br />
<li>Reporter Module</li><br />
<img src="https://static.igem.org/mediawiki/2014/8/8c/RepMod.JPG"><br />
<p align="justify">The construction of reporter module is consist of inducible promoter PibpAB <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K339010">(BBa_K339010)</a>, RBS <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, amilCP <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K592025">(BBa_K592025) </a>, and double terminator <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0015"> (Bba_B0015) </a>. BBa_K1387000 consist of RBS (Bba_B0034), amilCP (BBa_K592025) and double terminator (BBa_B0015), while the complete module was contained in <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K1387001">(Bba_K1387001)</a>. The reporter module will have a blue chromoprotein when the inclusion body is formed in the cell. This mechanism then acts as an indicator of cytoplasmic stress, in this situation is caused by inclussion body.</p><br />
<br><li>Degradation Module</li><br />
<img src="https://static.igem.org/mediawiki/2014/6/69/DegMod.JPG"><br />
<p align="justify">The casette of degradation module <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K1387006">(BBa_K1387006) </a> is comprised of constitutive promoter, tet operator, RBS, outer membrane protein A (ompA) fused with LC Cutinase, and double terminator. By using constitutive promoter, we expect the recombinant protein, in this case ompA-LC Cutinase, will be synthesized indefinitely. OmpA is outer membrane protein, it comprises of β–strand B3-B7. Rather than using full sequence of ompA, we use ompA (46-159) fused with lipoprotein signal sequence, thus exposing it on the cell surface of bacteria1. We fuse lpp-ompA with LC Cutinase, an enzyme that exhibit esterase activity. LC-Cutinase esterase activity will cleave the ester bond on PET and releasing terephthalic acid and ethylene glycol as a product. Fusion strategy of lpp-ompA on LC Cutinase gene is one of our way to create a whole cell biocatalyst which will be an effective molecular machinery to degrade PET on the environment. Due to constitutive expression of the recombinant protein in E.coli, there is a possibility of formation of insoluble protein inside the cell (inclusion body), because of that, we put tet operator upstream of RBS for further regulation (Georgiou et al, 1996).</p><br />
<br><li>Self-Regulatory Module</li><br />
<img src="https://static.igem.org/mediawiki/2014/b/b4/Sr1.JPG"><br />
<p align="justify">pIBPAB is a hybrid promoter that induced by inclusion body from unfunctional protein. This promoter followed by strong RBS <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, make a new part called <a style="color:#342D7E" href="http://parts.igem.org/Part:Bba_K1387005"> Bba_K1387005 </a> </p><br />
<img src="https://static.igem.org/mediawiki/2014/d/de/Sr2.JPG"><br />
<p align="justify">This casette <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0034">(BBa_K1387002)</a> consist of TetR <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_C0040">(BBa_C0040)</a> and Double Terminator <a style="color:#342D7E"href="http://parts.igem.org/Part:BBa_B0015"> (Bba_B0015) </a>. TetR is tetracycline repressor that bind to operator region and repressing the expression of downstream gene. In our system, the degradation module consist of TetO (Tet Operator) which is region of TetR (Tet Repressor) placed.</p><br />
<br><img src="https://static.igem.org/mediawiki/2014/f/f2/Sr3.JPG"><br />
<p>The construction of reporter module is consist of inducible promoter PibpAB <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K339010">(BBa_K339010)</a>RBS <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, amilCP <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K592025">(BBa_K592025) </a>, and double terminator <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0015"> (Bba_B0015) </a>. <a style="color:#342D7E"href="hhttp://parts.igem.org/Part:BBa_K1387000"> BBa_K1387000 </a> consist of RBS <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, amilCP <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K592025">(BBa_K592025) </a> and double terminator <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0015"> (Bba_B0015) </a>, while the complete module was contained in <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K1387001">BBa_K1387001 </a>. The reporter module will have a blue chromoprotein when the inclusion body is formed in the cell. This mechanism then acts as an indicator of cytoplasmic stress, in this situation is caused by inclussion body.</p><br />
<br><li>Convertion Module</li><br />
<img src="https://static.igem.org/mediawiki/2014/7/7a/ConMod.JPG"><br />
<p align="justify">Improvisation the part submitted by UC Davis team <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K936024">BBa_K936024</a> by combining the construct with terminator, so we can characterize the expression of the enzymes and performe the assay for ethylene glycol utilization better than before. We name this BioBrick as convertion module.</p><br />
</ol><br />
<br><li>References</li><br />
<p align="justify">Georgiou,G., Stephens,L., Stathopoulos,C., Poetschke,L., Mendenhall,J., and Earhart,F. 1996. Display of β-Lactamase on Eschericia coli Surface : Outer Membrane Phenotypes Conferred by Lpp’-ompA’- β-Lactamase Fusions. <i> Protein Engineering, 9, 239-247</i>.</p><br />
</ol><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Parts
Team:ITB Indonesia/Parts
2014-10-18T02:03:58Z
<p>Erawijantari: </p>
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>PARTS</h1><br />
<br><br />
<img src="https://static.igem.org/mediawiki/2014/d/db/Parts1.JPG"><br />
<br><br />
<br><br />
<h1>PARTS DESCRIPTION</h1><br />
<br><br />
<ol type="A"><br />
<li>Reporter Module</li><br />
<img src="https://static.igem.org/mediawiki/2014/8/8c/RepMod.JPG"><br />
<p align="justify">The construction of reporter module is consist of inducible promoter PibpAB <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K339010">(BBa_K339010)</a>, RBS <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, amilCP <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K592025">(BBa_K592025) </a>, and double terminator <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0015"> (Bba_B0015) </a>. BBa_K1387000 consist of RBS (Bba_B0034), amilCP (BBa_K592025) and double terminator (BBa_B0015), while the complete module was contained in <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K1387001">(Bba_K1387001)</a>. The reporter module will have a blue chromoprotein when the inclusion body is formed in the cell. This mechanism then acts as an indicator of cytoplasmic stress, in this situation is caused by inclussion body.</p><br />
<br><li>Degradation Module</li><br />
<img src="https://static.igem.org/mediawiki/2014/6/69/DegMod.JPG"><br />
<p align="justify">The casette of degradation module <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K1387006">(BBa_K1387006) </a> is comprised of constitutive promoter, tet operator, RBS, outer membrane protein A (ompA) fused with LC Cutinase, and double terminator. By using constitutive promoter, we expect the recombinant protein, in this case ompA-LC Cutinase, will be synthesized indefinitely. OmpA is outer membrane protein, it comprises of β–strand B3-B7. Rather than using full sequence of ompA, we use ompA (46-159) fused with lipoprotein signal sequence, thus exposing it on the cell surface of bacteria1. We fuse lpp-ompA with LC Cutinase, an enzyme that exhibit esterase activity. LC-Cutinase esterase activity will cleave the ester bond on PET and releasing terephthalic acid and ethylene glycol as a product. Fusion strategy of lpp-ompA on LC Cutinase gene is one of our way to create a whole cell biocatalyst which will be an effective molecular machinery to degrade PET on the environment. Due to constitutive expression of the recombinant protein in E.coli, there is a possibility of formation of insoluble protein inside the cell (inclusion body), because of that, we put tet operator upstream of RBS for further regulation (Georgiou et al, 1996).</p><br />
<br><li>Self-Regulatory Module</li><br />
<img src="https://static.igem.org/mediawiki/2014/b/b4/Sr1.JPG"><br />
<p align="justify">pIBPAB is a hybrid promoter that induced by inclusion body from unfunctional protein. This promoter followed by strong RBS <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, make a new part called <a style="color:#342D7E" href="http://parts.igem.org/Part:Bba_K1387005"> Bba_K1387005 </a> </p><br />
<img src="https://static.igem.org/mediawiki/2014/d/de/Sr2.JPG"><br />
<p align="justify">This casette <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0034">(BBa_K1387002)</a> consist of TetR <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_C0040">(BBa_C0040)</a> and Double Terminator <a style="color:#342D7E"href="http://parts.igem.org/Part:BBa_B0015"> (Bba_B0015) </a>. TetR is tetracycline repressor that bind to operator region and repressing the expression of downstream gene. In our system, the degradation module consist of TetO (Tet Operator) which is region of TetR (Tet Repressor) placed.</p><br />
<br><img src="https://static.igem.org/mediawiki/2014/f/f2/Sr3.JPG"><br />
<p>The construction of reporter module is consist of inducible promoter PibpAB <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K339010">(BBa_K339010)</a>RBS <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, amilCP <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K592025">(BBa_K592025) </a>, and double terminator <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0015"> (Bba_B0015) </a>. <a style="color:#342D7E"href="hhttp://parts.igem.org/Part:BBa_K1387000"> BBa_K1387000 </a> consist of RBS <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, amilCP <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K592025">(BBa_K592025) </a> and double terminator <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0015"> (Bba_B0015) </a>, while the complete module was contained in <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K1387001">BBa_K1387001 </a>. The reporter module will have a blue chromoprotein when the inclusion body is formed in the cell. This mechanism then acts as an indicator of cytoplasmic stress, in this situation is caused by inclussion body.</p><br />
<br><li>Convertion Module</li><br />
<img src="https://static.igem.org/mediawiki/2014/7/7a/ConMod.JPG"><br />
<p align="justify">Improvisation the part submitted by UC Davis team <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K936024">BBa_K936024</a> by combining the construct with terminator, so we can characterize the expression of the enzymes and performe the assay for ethylene glycol utilization better than before. We name this BioBrick as convertion module.</p><br />
<br><li>References</li><br />
<p>1Georgiou,G., Stephens,L., Stathopoulos,C., Poetschke,L., Mendenhall,J., and Earhart,F. 1996. Display of β-Lactamase on Eschericia coli Surface : Outer Membrane Phenotypes Conferred by Lpp’-ompA’- β-Lactamase Fusions. <i> Protein Engineering, 9, 239-247</i>.</p><br />
</ol><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Data
Team:ITB Indonesia/Data
2014-10-17T23:06:01Z
<p>Erawijantari: </p>
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<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
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</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>A. Scaning Electron Microscope (SEM) Result Analysis</h1><br />
<p align="justify">For biodegrading experiments, we used 3x5 cm <sup> 2 </sup> bottle plastics with average weight 0.05 g. Then, we washed it several times with water and ethanol.</p><br />
<p align="justify">Plastic samples were applied just before we inoculated the bacteria on the medium. Bacterial culture was grown at 37<sup>0</sup>C in Luria-Broth medium. After 3 days of incubation, we washed the plastics with water and ethanol then dried for measurement of the weight loss.</p><br />
<p align="justify">Meanwhile, for UC Davis bacterial culture, we inoculated the bacteria in Luria-Broth medium supplemented with 170 ppm cloramphenicol. After an absorbance A600 nm of 0.6 was reached, we added 1% of arabinose and plastic samples. After 2 days of incubation, we washed the plastics with water and ethanol then dried for measurement of the weight loss.</p><br />
<p align="justify">We used medium supplemented with antibiotics and plastics sample as control on this experiment. The scaning electron microsccopy analysis of fractured surface of PET film was carried out using Scaning Electron Microscope. The surface of the treated PET samples were coated with conductive heavy metals such as gold/palladium.</p><br />
<palign="justify">SEM image shows that cracks were observed at the surface of a plastic samples (PET) after incubation of the bacterial culture. However, there was no significant weight decreased of the plastics samples. From this SEM study we concluded that both our LC Cut UC Davis and OmpA-LC-Cut Fussion ITB 2014 able to degrade PET plastics samples. Figure 1. shown the control sampel that we could not observe any cracks. Figure 2 and 3 shown the PET degradation by Lc cutinase from UC davis and from ITB_Indonesia.</p><br />
<br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/e/e9/Sem1.JPG"><br />
<p><small>Figure 1. Control sampel (PET plastic incubated in medium)</small></p><br />
</div><br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/9/92/Sem2.JPG"><br />
<p><small>Figure 2. PET plastic after treatment with ompA-LC-cutinase from ITB_Indonesia construct (BBa_K1387006)</small></p><br />
</div><br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/6/62/Sem3.JPG"><br />
<p><small>Figure 3. PET plastic after treatment with LC-cutinase UC Davis 2012 gene construct (Bba_K936020)</small></p><br />
</div><br />
<br />
<h3>References</h3><br />
<ol style="text-decoration:none;"><br />
<li>Sepperumal, Umaheswari, Murali Markandan, and Anbusaravanan Natarajan. 2013. electron microscopic studies of Polyethylene terepthalate degradation potential of Pseudomonas species. J. Microbiol. Biotech. Res. (1): 104-110</li><br />
<li>Mittal,Alok, R.K. Soni, Khrisna Dutt, and Swati Singh. 2010. Scaning electron microscopy study of hazardous waste flakes of polyethylene terephthalate (PET) by aminolysis and ammonolysis. Journal of Hazardous Materials Volume 178, Issues 1-3 </li><br />
</ol><br />
<br />
<br><br />
<h1>B. Ethylene Glikol Chromic Acid Assay</h1><br />
<p align="justify">Poly(ethylene terephthalate) (PET) is a plastic that is a polymer of ethylene terephthalate (C10H8O4) units (Sarker et al., 2010). Polyethylene terephthalate (PET) can be degrade via limited enzymatic hydrolysis of the ester bond of the polymer backbone, one enzyme that have been assesed for this purpose is cutinase (Ribitsch et al., 2012). When polyethylene terephthalate degraded, it will produce terephthalic acid and ethylene glycol (Venkatachalam et al., 2012). Ethylene glycol is one of alcohol compound. Oxidation of alcohols by strong oxidants such as chromic acid (K2Cr2O7) in suphuric acid(H2SO4) is possible, but differs depending on the degree of alcohol. If a reaction has occurred using chromic acid in sulphuric acid, there is a color change from orange to green. In our project we use chromic acid to detect ethylene glycol as product of the PET degradation by LC cutinase enzyme.</p><br />
<br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/2/2e/Sem5.JPG"><br />
</div><br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/2/23/Sem4.JPG"><br />
</div><br />
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<p align="justify">Each sample used for measure the absorbance contain a fixed amount of chromic acid. The absorbance of the chromic acid will represent the total amount of ethylene glycol inversely. It means that if the absorbance is high, the amount of ethylene glycol in solution is low, and if the absorbance is low, the amount of ethylene glycol in solution is high. If the absorbance is high, the amount of chromic acid is high, which means that the amount of ethylene glycol being oxidized by chromic acid is low and vice versa. Both of LC Cutinase UC Davis and OmpA-LC Cutinase ITB_Indonesia shown the degrading activity of PET become ethylene glycol as once of the product.The optimum time to produce ethylene glycol is on the fourth hour after inoculation.</p><br />
<br />
<h3>References</h3><br />
<ol style="text-decoration:none;"><br />
<li>Ribitsch D, Enrique HA, Katrin G, Anita D, Sabine Z, Annemarie M, Rosario DR, Georg S, Karl G, Helmut S and Georg MG. 2012. A New Esterase from Thermobifida halotolerans Hydrolyses Polyethylene Terephthalate (PET) and Polylactic Acid (PLA). <i>Polymers. 4: 617-629</i></li><br />
<li>Sarker M, Aminul K, Mohammad MR, Mohammed M and ASMD Mohammad. 2011. Waste Polyethylene Terephthalate (PETE-1) Conversion into Liquid Fuel. <i>Journal of Fundamentals of Renewable Energy and Applications. 1</i></li><br />
<li>Venkatachalam S, Shilpa GN, Jayprakash VL, Prashant RG, Krishna R and Anil KK. 2012. Degradation and Recyclability of Poly (Ethylene Terephthalate). <i>Intech</i></li><br />
</ol><br />
<br />
<br><br />
<h1>C. pNPP assay</h1><br />
<p align="justify">Blastp program shows that LC cutinase shows high amino acid sequence similarity (54 to 60%) to lipase (Sulaiman, 2012). So, we tried to measure LC-cutinase activity using p-nitrophenylpalmitate (pNPP) as a substrate. pNPP typically used for measuring lipase activity.</p><br />
<p>In this assay, we used bacterial culture because in real application we would like to use bacterial culture to degrade PET directly. As negative control, we used <i>''E.coli'' </i> BL21(DE3) without plasmid. We also included LC-cutinase UC Davis 2012 gene construct (Bba_K936020) in this experiment. LC-cutinase UC Davis 2012 bacterial culture is induced after 2 hours of growth (after OD 600 nm ±0.6 was reached) using 1% of arabinose. All of the bacterial culture were harvested after 4 hours of growth.From ethylene glycol assay using chromic acid, we found that ompA-LC-cutinase from ITB_Indonesia construct (BBa_K1387006) has the highest activity after 4 hours of growth.</p><br />
<p align="justify">At first, we measured the activity at 60 degrees Celsius. We found that both ompA-LC-cutinase from ITB_Indonesia and LC-cutinase UC Davis 2012 still performed small activity. Then, we tried measure the activity at 25 degrees Celsius. The ompA-LC-cutinase from ITB_Indonesia’s activity was 0.001 U/mL. While, LC-cutinase UC Davis 2012 has ten fold higher (0.01 U/mL). The LC cutinase activity was determined based on the standard curve of p-nitrophenol. One unit of cutinase activity was defined as the amount of enzyme releasing 1 μmol pNP per minute under the assay conditions.</p><br />
<p align="justify">From this experiment, we are able to confirm that ompA-LC-cutinase from ITB_Indonesia successfully perform activity. But, its suggest that we still have to check the possible activity in different time of bacterial growth, expression rate, LC-cutinase presentation by ompA and activity assay using different substrate.</p><br />
<br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/9/90/Sem6.JPG"><br />
<p><small>Figure 1. LC Cutinase activity. The enzymatic activity was determined at 60C in assay condition using pNP-palmitate (C16) as a substrate. The experiment was carried out twice.</small></p><br />
</div><br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/a/a1/Sem7.JPG"><br />
<p><small>Figure 2. LC Cutinase activity. The enzymatic activity was determined at 25 C in assay condition using pNP-palmitate (C16) as a substrate. The experiment was carried out twice.</small></p><br />
</div><br />
<p align"justify">Small activity of LC-Cutinase maybe due to their oligomeric state. Tethering protein to cell surface may affect folding of protein and also their biological function (Park,2010). These suggest us to look futher detail on folding and displaying state of LC cutinase.</p><br />
<br />
<p align="justify">There are several things that should be noted when we designate fusion strategy of passanger protein or protein of interest with the outer membrane of protein. Linking the different passanger proteins to the same outer membrane protein may result in different translocation. The characteristic of passanger protein affects the translocation. Its characteristic such as formation of disulfide bridge, total residue of hydrophobic aminoacid etc. </p><br />
<p align="justify">So there are four requirements have to be met for constructing well designed surface display. First is, it has to possesses efficient signal peptide. Second, it should have strong anchor to keep the fusion protein from detachment. Third, it should be compatible to the sequence that is inserted or fused. The last is it should resisstant to the attack of the protease from medium or periplasmic membrane. </p><br />
<br />
<h3>Reference</h3><br />
<ol style="text-decoration:none;"><br />
<li>Sulaiman, Sintawee, SayaYamat, EikoKanaya, Joong-Jae Kim, Yuichi Koga, Kazufumi Takano and Shigenori Kanaya. 2012. Isolation of a Novel Cutinase Homolog with Polyethylene Terephthalate-Degrading Activity from Leaf-Branch Compost by Using a Metagenomic Approach. <i>Applied and Environmental Microbiology p 1556-1562.</i></li><br />
<li>Lee, Sang Yup., Choi, Jong Hyun., Xu, Zhaohui. 2003. “Microbial Cell-Surface Display” <i>Elsevier</i></li><br />
</ol><br />
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2014-10-17T23:02:49Z
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<p align="justify">This project is done by iGEM ITB_Indonesia 2014 team members and our collaborator . Our team have worked well dan dedicated for this project. </p><br />
<h1>Project name: SynBIGreen (Synthetic Biology Indonesia Go Green)</h1><br />
<p align="justify">Our team consist of variant background of knowledge, they are from science and engineering. We have many talents that no have one for the others. Each member are important and have different roles and responsibilities that can lead us to reach our target. For finishing this project we mapped our talent to variant job following as:</p><br />
<br />
<h3>Team Leader :</h3><br />
<p>Joko Pebrianto Trinugroho</p><br />
<br />
<h3>Construct :</h3><br />
<p>Tri Ekawati Heryanto, Pande Putu Erawijantari, Oktira Roka Aji, Joko Pebrianto Trinugroho, Nimas Ghassani</p><br />
<br />
<h3>Modeling System :</h3><br />
<p>Aditya Putra Pratama</p><br />
<br />
<h3>Sponsorship Team :</h3><br />
<p>Lance Rosa Karo-karo, Oktira Roka Aji, Kenia Permata Sukma, Wiyudi Gomulya</p><br />
<br />
<h3>Visual Graphic Design Team :</h3><br />
<p>Fania Feby Ramadhani</p><br />
<br />
<h3>Wiki :</h3><br />
<p>Muhammad Hariomurti Mardikusumo, Rizky Kusumah</p><br />
<br />
<h3>Human practice team :</h3><br />
<p>Nimas Ghasani, Risma Wiharyanti, Wuddan Nadhirah Rodiana, Bakhtiar Hermawan</p><br />
<br />
<h3>Public Relation :</h3><br />
<p>Mardalisa</p><br />
<br />
<h3>Project Idea :</h3><br />
<p>Tirta Widi Gilang Citradi, Yovin, Christian Heryakusuma</p><br />
<br />
<h3>General Support :</h3><br />
<p>Dr. Sony Suhandono, Dr. Dessy Natalia, Dr. Maelita R. Moeis helped us during our brainstorming<br>Dr. Dessy Natalia is letting us use her lab for some experiments</p><br />
<br />
<h3>Modeling Support :</h3><br />
<p>Melia Silmi and Faisal Rahmananda are bachelor students , department of mathematics help us to model our system<br>Dr. Mochamad Apri is our Instructor and lecturer in department of mathematics. He is an expert on mathematical modelling and helped our team to model our system.</p><br />
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<h3>Imaging Support :</h3><br />
<p>Faculty of Mathematical and Natural Science Institut Teknologi Bandung helped with the Scanning Electron Microscope (SEM) analysis to our degradation system</p><br />
<br />
<h3>Video Support:</h3><br />
<p> Muhammad Daniel Septian, magister student majoring design in Faculty of Arts and Design help us to make a project video</p><br />
<br />
<h3>Wiki Support</h3><br />
<p>Riandy Rahman Nugraha is a bachelor students, department of Informatics Engineering guided us to make our wiki</p> <br />
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Erawijantari
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Team:ITB Indonesia/Attributions
2014-10-17T23:01:37Z
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<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<p align="justify">This project is done by iGEM ITB_Indonesia 2014 team members and our collaborator . Our team have worked well dan dedicated for this project. </p><br />
<h1>Project name: SynBIGreen (Synthetic Biology Indonesia Go Green)</h1><br />
<p align="justify">Our team consist of variant background of knowledge, they are from science and engineering. We have many talents that no have one for the others. Each member are important and have different roles and responsibilities that can lead us to reach our target. For finishing this project we mapped our talent to variant job following as:</p><br />
<br />
<h3>Team Leader :</h3><br />
<p>Joko Pebrianto Trinugroho</p><br />
<br />
<h3>Construct :</h3><br />
<p>Tri Ekawati Heryanto, Pande Putu Erawijantari, Oktira Roka Aji, Joko Pebrianto Trinugroho, Nimas Ghassani</p><br />
<br />
<h3>Modeling System :</h3><br />
<p>Aditya Putra Pratama</p><br />
<br />
<h3>Sponsorship team :</h3><br />
<p>Lance Rosa Karo-karo, Oktira Roka Aji, Kenia Permata Sukma, Wiyudi Gomulya</p><br />
<br />
<h3>Visual graphic design team :</h3><br />
<p>Fania Feby Ramadhani</p><br />
<br />
<h3>Wiki :</h3><br />
<p>Muhammad Hariomurti Mardikusumo, Rizky Kusumah</p><br />
<br />
<h3>Human practice team :</h3><br />
<p>Nimas Ghasani, Risma Wiharyanti, Wuddan Nadhirah Rodiana, Bakhtiar Hermawan</p><br />
<br />
<h3>Public Relation :</h3><br />
<p>Mardalisa</p><br />
<br />
<h3>Project Idea :</h3><br />
<p>Tirta Widi Gilang Citradi, Yovin, Christian Heryakusuma</p><br />
<br />
<h3>General Support :</h3><br />
<p>Dr. Sony Suhandono, Dr. Dessy Natalia, Dr. Maelita R. Moeis helped us during our brainstorming<br>Dr. Dessy Natalia is letting us use her lab for some experiments</p><br />
<br />
<h3>Modeling Support :</h3><br />
<p>Melia Silmi and Faisal Rahmananda are bachelor students , department of mathematics help us to model our system<br>Dr. Mochamad Apri is our Instructor and lecturer in department of mathematics. He is an expert on mathematical modelling and helped our team to model our system.</p><br />
<br />
<h3>Imaging Support :</h3><br />
<p>Faculty of Mathematical and Natural Science Institut Teknologi Bandung helped with the Scanning Electron Microscope (SEM) analysis to our degradation system</p><br />
<br />
<h3>Video Support:</h3><br />
<p> Muhammad Daniel Septian, magister student majoring design in Faculty of Arts and Design help us to make a project video</p><br />
<br />
<h3>Wiki Support</h3><br />
<p>Riandy Rahman Nugraha is a bachelor students, department of Informatics Engineering guided us to make our wiki</p> <br />
<br />
<br><br />
<br><br />
<br />
<div id="sponsor"></div><br />
</div><br />
</div><br />
</body><br />
</html></div>
Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Medsos
Team:ITB Indonesia/Medsos
2014-10-17T16:53:00Z
<p>Erawijantari: </p>
<hr />
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<br />
<br />
</style><br />
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</head><br />
<body><br />
<div id="putih"> <div id="logo-igem" onclick="window.location.href='https://2014.igem.org/'"></div><br />
<div id="center"><br />
<div id="logo"></div><br />
<br />
<div id="header"></div><br />
<br />
<div id="blockNav"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Social Media</h1><br />
<p style="text-align: justify">In this digital era, internet users highly increase. Social media is an effective tool that can be used to introduce synthetic biology, iGEM, and ITB_Indonesia team. We frequently used Facebook and twitter as our link to society because these social media are the most popular media in Indonesia with the highest access number.</p><br />
<p style="text-align: justify">Today, internet users in Indonesia has reached 82 million people and still growing with very wide range age users, from kids up to adult. Besides, Indonesia is placed on the 8th position in the world as the highest internet users and holds the 5th position as the most Twitter users with 29,000 twitter account. It is an opportunity for ITB_Indonesia team to introduce iGEM and synthetic biology. With these facilities, ITB_Indonesia team hopes that people can access iGEM and synthetic biology easily.</p><br />
<p style="text-align: justify">ITB_Indonesia team has a Facebook fan page with 1,474 likes and 151 followers on twitter. Here, we introduce our team, idea, project, and share our activity. We also tell information about synthetic biology generally.</p><br><br />
<br />
<table><br />
<tr><br />
<td><a href="https://www.facebook.com/synbio.itb/info"><img src="https://static.igem.org/mediawiki/2014/c/ca/Fanpage_fb.jpg"></a></td><br />
<td style="width:60px"></td><br />
<td><a href="https://twitter.com/synbio_itb"><img src="https://static.igem.org/mediawiki/2014/0/05/Twit.JPG"></a></td><br />
</tr><br />
</table><br />
<br />
<br />
<br><br />
<br><br />
<br />
<div id="sponsor"></div><br />
</div><br />
</div><br />
</body><br />
</html></div>
Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/nb-wetlab
Team:ITB Indonesia/nb-wetlab
2014-10-17T16:40:05Z
<p>Erawijantari: </p>
<hr />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
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<br />
<br><br />
<h1><center>June</center></h1><br />
<h3>Week 1</h3><br />
<p align="justify">In the first week of June we observed the laboratory. We made a list of materials that we need for future wetlab activities. We prepared LB medium, searched the antibiotic optimization method and made competent cells using CCMB 80 buffer. We also sterilized all of the equipment needed. We already decided our idea that is engineer bacteria that can degrade plastic. Previously, we had four main ideas:<br><br />
1. Microalgae biofuel<br>2. Synthetic bacteria that can degrade plastic<br>3. Urine bio-sensor<br>4. Cyanide bio-sensor<br><br />
After competent cells were ready and antibiotic optimization had done then we had plan to try re-hydrated some parts and transform it to our <i> Escherichia coli </i> DH5α competent cells. We also had decided our construct so we would ready to order some synthetic parts and ordered it to iGEM HQ especially for some parts that not provided in the 2014 parts.</p><br />
<h3>Week 2</h3><br />
<p align="justify">This was our first time to see the iGEM distribution kit plate which was so exciting. We would make a trial to transform iGEM 2013 distribution kit using our competent cells. That transformation was not good on first, second, and third trial. Then we concluded that we must try the iGEM distribution kit plate 2014. We also tried to test our competent cells efficiency and we did not face any problem with our competent cells.</p><br />
<h3>Week 3</h3><br />
<p align="justify">Finally, we re-hydrated parts from iGEM kit plate 2014 which had many copies to make sure our method was optimal enough to transform those iGEM parts. We had already a plan to make four systems for our bacteria to degrade plastic. We divided our team into four groups. Each groups focused to each systems. We tried to transform BBa_K592025, BBa_B0015, BBa_B0034, BBa_I0040. The transformation did not have any validate colonies.</p><br />
<h3>Week 4</h3><br />
<p align="justify">We targeted to transform parts from iGEM distribution kit. Finally, we could see our red colony from RFP parts. So, we made liquid culture and wait until 16 hours. Then the medium turned into red. We tried to isolate the plasmid using Alkaline Lysis Method. The confirmation of the plasmids isolated were by electrophoresis, restriction and PCR method. The result was positive.</p><br />
<br><br />
<br />
<h1><center>July</center></h1><br />
<h3>Week 1</h3><br />
<p>Finnaly our constructs are already fixed. We ordered our synthetic gene for our main construct and ordered some parts from iGEM Headquarters. Our competent cell was also ready for transforming the other parts that we will use for our construct. We rehydrated BBa_K592025, BBa_J04450, BBa_C0040, BBa_B0017 and BBa_B0015, the colonies was growing well and ready for plasmid isolation. All of the plasmid was confirmed using restriction method and the result was positive. In this first week of July we learnt more about 3A assembly since we are ready to make our construct as well.</p><br />
<h3>Week 2</h3><br />
<p>We are ready to construct our building blocks. But first, we made some optimization method for double digest restriction reaction including for time incubation and reaction compotition. We also received our parts that we had ordered from iGEM headquarter BBa_K103006, BBa_K228004, BBa_K936024, BBa_K339010. We also tried to build our construct using two methods: standard assembly and 3A assembly. Still not work well. In this week we also elaborated and had brainstorming for our next method after our construct had been done.</p><br />
<h3>Week 3</h3><br />
<p>We were ready to try 3A assembly again, we made four different ligation composition to make sure which one will work well. After transformation, finally we got few colony in our petri disc, ready for plasmid isolation. After confirmation for all of the plasmid, we got no right colony yet. We then tried again using PSB1C3 with Rfp for the plasmid backbone instead of linearized plasmid to make it easier for colony selection using red-white screening. Still not got our right construct. Let the challenge begin and let’s do the best for it.</p><br />
<h3>Week 4</h3><br />
<p>Challenge accepted, we did our 3A again. We discussed about our failure for the 3A this two weeks and got some conclusion. Finally for this trial we got some colony which was the right construct BBa_K1387002. We then found the optimum ligation composition and reaction. Ready for another construct for reporter and converter module while waiting for synthetic gene for our main construct.</p><br />
<br><br />
<br />
<h1><center>August</center></h1><br />
<h3>Week 1</h3><br />
<p align="justify">Luckily, the primers were arrived. We ordered universal and spesific primer to check our construct using PCR technique. For optimization, we used gradient PCR technique. By using this technique, we were able to get a good results. Unfortunately, many of our construct were still in pSB1K3 backbone, so we must translocate it to pSB1C3 backbone.</p><br />
<h3>Week 2</h3><br />
<p align="justify">In this week we are trying to make our 3A assembly again since we still have some gene construct that must be finished before our synthetic gene comes. From the 3A assembly we got several colony that ready for plasmid isolation. After the confirmation through PCR methode, we did not get the right colony yet, so we must try the other compotition in ligation compotition, because we did not face any problem in transformation. After trying any ligation reaction , we got the optimal compotition for some construct, but the other still become our big mystery. In this week, we are also want to try the construct from UC Davis for Lc Cutinase BBa_K936020 and ethylene glycol converter module BBa_K936024. We did plasmid isolation from that part and transform it to the E.coli BL21 (DE3) We try to test the gene expression and test the product through SDS-PAGE method and we can see the protein band from both of the construct but not as the thick band. We also try to sequencing the positive construct in this week.</p><br />
<h3>Week 3</h3><br />
<p align="justify">Our target in this third week of august is to finish all of the construct except the construct that need synthetic gene due the parts submission deadline is coming. Restriction, purification gell (if needed), ligation, transformation, plasmid isolation, and confirmation is become our routine activities. In this week, we also learn more about how to express our protein, how to test our construct, how to measure the construct activities and did SDS-PAGE analysis.</p><br />
<h3>Week 4</h3><br />
<p align="justify">We focused to make a simple trial for our method to test the protein band of cutinase, and conversion methode, and discuss about our construct characterization especially for OmpA-LcCutinase construct (PET degradation module) as our main construct. We also discuss for synthetic gene cloning method. In this week we also focused on finishing our construct in PSB1C3 backbone and ready for shipment. Brainstoarming also become our routine activities because we found some failure in this week and feel not ready yet facing the parts submision deadline. We also try to elaborate how to make a modeling for our system and discuss with the expert.</p><br />
<br><br />
<br />
<h1><center>September</center></h1><br />
<h3>Week 1</h3><br />
<p align="justify">We have to prepare our parts submission form and check all of positive construct to make sure the condition of the construct is still good. In this week, synthetic gene, the thing that we waiting for one and half month finnaly arrived in our campuss. We try to make optimal reaction for our limited stock of synthetic gene. We try to clone our synthetic gene into some backbone including some of commercial clone vector and PSB1C3 of course. No collonies observed, so desperate and we discuss and evaluate our method and take a note of some mistake that we did in this method. Fortunatelly we have primer for our synthetic gene so we could do amplification using PCR for our synthetic gene. We also make new competent cell and trying electhrophoration transformation method instead of common heat shock methode. Still, no colonies observed.</p><br />
<h3>Week 2</h3><br />
<p align="justify">We try to clone our synthetic again. We must be succes in this week to step forward finishing our main construct. We try several method and try to use overnight ligation method. Finnaly, we got the positive colony, so happy. We also checked our plasmid again and found that some of our construct disappear but the other is still in good condition. We also make a optimization for PCR using touchdown and temperature gradient method to check the good temperature condition for the primer.</p><br />
<h3>Week 3</h3><br />
<p align="justify">We did PCR for construct confirmation for the collonies through crude PCR method using the optimal annealing temperature. We also did plasmid isolation for the positive colonies. We also make the method plan for PET degradation characterization using our construct and compared to UC Davis construct. We make some method for measurement including pH detection, ethylene glycol detection using dichromic acid method, and check the correlation between cell number in PET degradation rate.</p><br />
<h3>Week 4</h3><br />
<p align="justify">We submitted some of our parts including BBa_K1387000, BBa_K1387002 and BBa_K1387006 In the 11 days remaining for parts submission, we still try to build our construct and start to think the future development of our system. We also try to help Brawijaya team to cloning some of their parts and synthetic gene. We will start our parts characterization using several method including ethylene glycol assay using chromatic acid, pNPP assay to check the activity, growth curve, SEM, and qualitative assay using tributirin agar method.</p><br />
<br><br />
<br />
<h1><center>October</center></h1><br />
<h3>Week 1</h3><br />
<p align="justify">The sending parts week for second term. three parts is ready to send including BBa_K1387001, BBa_K1387006 and BBa_K1387005. Our dry lab team is also ready for modelling. We got some parameter to make the modelling. We also got some idea for our future system.</p><br />
<h3>Week 2</h3><br />
<p align="justify"> This is our busiest week for iGEM because of many upcoming deadline. This week we constructed our parts that still not complete yet. We also did characterization for our main parts including pNPP assay, ethylene glicol assay using chromic acid, and SEM analysis. We compared our LC-cutinase gene construct with cutinase gene construct for UC Davis 2012 using our characterization method. This week we got all of characterization data and also sent our second batch for parts submission due to the deadline of parts submission (Bba_K1387006, Bba_K1387001, and Bba_K1387005).</p><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/nb-wetlab
Team:ITB Indonesia/nb-wetlab
2014-10-17T16:26:30Z
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
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<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
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<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1><center>June</center></h1><br />
<h3>Week 1</h3><br />
<p align="justify">In the first week of June we observed the laboratory. We made a list of materials that we need for future wetlab activities. We prepared LB medium, searched the antibiotic optimization method and made competent cells using CCMB 80 buffer. We also sterilized all of the equipment needed. We already decided our idea that is engineer bacteria that can degrade plastic. Previously, we had four main ideas:<br><br />
1. Microalgae biofuel<br>2. Synthetic bacteria that can degrade plastic<br>3. Urine bio-sensor<br>4. Cyanide bio-sensor<br><br />
After competent cells were ready and antibiotic optimization had done then we had plan to try re-hydrated some parts and transform it to our <i> Escherichia coli </i> DH5α competent cells. We also had decided our construct so we would ready to order some synthetic parts and ordered it to iGEM HQ especially for some parts that not provided in the 2014 parts.</p><br />
<h3>Week 2</h3><br />
<p align="justify">This was our first time to see the iGEM distribution kit plate which was so exciting. We would make a trial to transform iGEM 2013 distribution kit using our competent cells. That transformation was not good on first, second, and third trial. Then we concluded that we must try the iGEM distribution kit plate 2014. We also tried to test our competent cells efficiency and we did not face any problem with our competent cells.</p><br />
<h3>Week 3</h3><br />
<p align="justify">Finally, we re-hydrated parts from iGEM kit plate 2014 which had many copies to make sure our method was optimal enough to transform those iGEM parts. We had already a plan to make four systems for our bacteria to degrade plastic. We divided our team into four groups. Each groups focused to each systems. We tried to transform BBa_K592025, BBa_B0015, BBa_B0034, BBa_I0040. The transformation did not have any validate colonies.</p><br />
<h3>Week 4</h3><br />
<p align="justify">We targeted to transform parts from iGEM distribution kit. Finally, we could see our red colony from RFP parts. So, we made liquid culture and wait until 16 hours. Then the medium turned into red. We tried to isolate the plasmid using Alkaline Lysis Method. The confirmation of the plasmids isolated were by electrophoresis, restriction and PCR method. The result was positive.</p><br />
<br><br />
<br />
<h1><center>July</center></h1><br />
<h3>Week 1</h3><br />
<p>Finnaly our constructs are already fixed. We ordered our synthetic gene for our main construct and ordered some parts from iGEM Headquarters. Our competent cell was also ready for transforming the other parts that we will use for our construct. We rehydrated BBa_K592025, BBa_J04450, BBa_C0040, BBa_B0017 and BBa_B0015, the colonies was growing well and ready for plasmid isolation. All of the plasmid was confirmed using restriction method and the result was positive. In this first week of July we learnt more about 3A assembly since we are ready to make our construct as well.</p><br />
<h3>Week 2</h3><br />
<p>We are ready to construct our building blocks. But first, we made some optimization method for double digest restriction reaction including for time incubation and reaction compotition. We also received our parts that we had ordered from iGEM headquarter BBa_K103006, BBa_K228004, BBa_K936024, BBa_K339010. We also tried to build our construct using two methods: standard assembly and 3A assembly. Still not work well. In this week we also elaborated and had brainstorming for our next method after our construct had been done.</p><br />
<h3>Week 3</h3><br />
<p>We were ready to try 3A assembly again, we made four different ligation composition to make sure which one will work well. After transformation, finally we got few colony in our petri disc, ready for plasmid isolation. After confirmation for all of the plasmid, we got no right colony yet. We then tried again using PSB1C3 with Rfp for the plasmid backbone instead of linearized plasmid to make it easier for colony selection using red-white screening. Still not got our right construct. Let the challenge begin and let’s do the best for it.</p><br />
<h3>Week 4</h3><br />
<p>Challenge accepted, we did our 3A again. We discussed about our failure for the 3A this two weeks and got some conclusion. Finally for this trial we got some colony which was the right construct BBa_K1387002. We then found the optimum ligation composition and reaction. Ready for another construct for reporter and converter module while waiting for synthetic gene for our main construct.</p><br />
<br><br />
<br />
<h1><center>August</center></h1><br />
<h3>Week 1</h3><br />
<p align="justify">Luckily, the primers were arrived. We ordered universal and spesific primer to check our construct using PCR technique. For optimization, we used gradient PCR technique. By using this technique, we were able to get a good results. Unfortunately, many of our construct were still in pSB1K3 backbone, so we must translocate it to pSB1C3 backbone.</p><br />
<h3>Week 2</h3><br />
<p align="justify">In this week we are trying to make our 3A assembly again since we still have some gene construct that must be finished before our synthetic gene comes. From the 3A assembly we got several colony that ready for plasmid isolation. After the confirmation through PCR methode, we did not get the right colony yet, so we must try the other compotition in ligation compotition, because we did not face any problem in transformation. After trying any ligation reaction , we got the optimal compotition for some construct, but the other still become our big mystery. In this week, we are also want to try the construct from UC Davis for Lc Cutinase BBa_K936020 and ethylene glycol converter module BBa_K936024. We did plasmid isolation from that part and transform it to the E.coli BL21 (DE3) We try to test the gene expression and test the product through SDS-PAGE method and we can see the protein band from both of the construct but not as the thick band. We also try to sequencing the positive construct in this week.</p><br />
<h3>Week 3</h3><br />
<p align="justify">Our target in this third week of august is to finish all of the construct except the construct that need synthetic gene due the parts submission deadline is coming. Restriction, purification gell (if needed), ligation, transformation, plasmid isolation, and confirmation is become our routine activities. In this week, we also learn more about how to express our protein, how to test our construct, how to measure the construct activities and did SDS-PAGE analysis.</p><br />
<h3>Week 4</h3><br />
<p align="justify">We focused to make a simple trial for our method to test the protein band of cutinase, and conversion methode, and discuss about our construct characterization especially for OmpA-LcCutinase construct (PET degradation module) as our main construct. We also discuss for synthetic gene cloning method. In this week we also focused on finishing our construct in PSB1C3 backbone and ready for shipment. Brainstoarming also become our routine activities because we found some failure in this week and feel not ready yet facing the parts submision deadline. We also try to elaborate how to make a modeling for our system and discuss with the expert.</p><br />
<br><br />
<br />
<h1><center>September</center></h1><br />
<h3>Week 1</h3><br />
<p align="justify">We have to prepare our parts submission form and check all of positive construct to make sure the condition of the construct is still good. In this week, synthetic gene, the thing that we waiting for one and half month finnaly arrived in our campuss. We try to make optimal reaction for our limited stock of synthetic gene. We try to clone our synthetic gene into some backbone including some of commercial clone vector and PSB1C3 of course. No collonies observed, so desperate and we discuss and evaluate our method and take a note of some mistake that we did in this method. Fortunatelly we have primer for our synthetic gene so we could do amplification using PCR for our synthetic gene. We also make new competent cell and trying electhrophoration transformation method instead of common heat shock methode. Still, no colonies observed.</p><br />
<h3>Week 2</h3><br />
<p align="justify">We try to clone our synthetic again. We must be succes in this week to step forward finishing our main construct. We try several method and try to use overnight ligation method. Finnaly, we got the positive colony, so happy. We also checked our plasmid again and found that some of our construct disappear but the other is still in good condition. We also make a optimization for PCR using touchdown and temperature gradient method to check the good temperature condition for the primer.</p><br />
<h3>Week 3</h3><br />
<p align="justify">We did PCR for construct confirmation for the collonies through crude PCR method using the optimal annealing temperature. We also did plasmid isolation for the positive colonies. We also make the method plan for PET degradation characterization using our construct and compared to UC Davis construct. We make some method for measurement including pH detection, ethylene glycol detection using dichromic acid method, and check the correlation between cell number in PET degradation rate.</p><br />
<h3>Week 4</h3><br />
<p align="justify">We submitted some of our parts including BBa_K1387000, BBa_K1387002 and BBa_K1387006 In the 11 days remaining for parts submission, we still try to build our construct and start to think the future development of our system. We also try to help Brawijaya team to cloning some of their parts and synthetic gene. We will start our parts characterization using several method including ethylene glycol assay using chromatic acid, pNPP assay to check the activity, growth curve, SEM, and qualitative assay using tributirin agar method.</p><br />
<br><br />
<br />
<h1><center>October</center></h1><br />
<h3>Week 1</h3><br />
<p align="justify">The sending parts week for second term. three parts is ready to send including BBa_K1387001, BBa_K1387006 and BBa_K1387005. Our dry lab team is also ready for modelling. We got some parameter to make the modelling. We also got some idea for our future system.</p><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/nb-wetlab
Team:ITB Indonesia/nb-wetlab
2014-10-17T16:25:40Z
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
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<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
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<li>PROJECT<br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
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<li>HUMAN PRACTICE<br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
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<br><br />
<h1><center>June</center></h1><br />
<h3>Week 1</h3><br />
<p align="justify">In the first week of June we observed the laboratory. We made a list of materials that we need for future wetlab activities. We prepared LB medium, searched the antibiotic optimization method and made competent cells using CCMB 80 buffer. We also sterilized all of the equipment needed. We already decided our idea that is engineer bacteria that can degrade plastic. Previously, we had four main ideas:<br><br />
1. Microalgae biofuel<br>2. Synthetic bacteria that can degrade plastic<br>3. Urine bio-sensor<br>4. Cyanide bio-sensor<br><br />
After competent cells were ready and antibiotic optimization had done then we had plan to try re-hydrated some parts and transform it to our <i> Escherichia coli </i> DH5α competent cells. We also had decided our construct so we would ready to order some synthetic parts and ordered it to iGEM HQ especially for some parts that not provided in the 2014 parts.</p><br />
<h3>Week 2</h3><br />
<p align="justify">This was our first time to see the iGEM distribution kit plate which was so exciting. We would make a trial to transform iGEM 2013 distribution kit using our competent cells. That transformation was not good on first, second, and third trial. Then we concluded that we must try the iGEM distribution kit plate 2014. We also tried to test our competent cells efficiency and we did not face any problem with our competent cells.</p><br />
<h3>Week 3</h3><br />
<p align="justify">Finally, we re-hydrated parts from iGEM kit plate 2014 which had many copies to make sure our method was optimal enough to transform those iGEM parts. We had already a plan to make four systems for our bacteria to degrade plastic. We divided our team into four groups. Each groups focused to each systems. We tried to transform BBa_K592025, BBa_B0015, BBa_B0034, BBa_I0040. The transformation did not have any validate colonies.</p><br />
<h3>Week 4</h3><br />
<p align="justify">We targeted to transform parts from iGEM distribution kit. Finally, we could see our red colony from RFP parts. So, we made liquid culture and wait until 16 hours. Then the medium turned into red. We tried to isolate the plasmid using Alkaline Lysis Method. The confirmation of the plasmids isolated were by electrophoresis, restriction and PCR method. The result was positive.</p><br />
<br><br />
<br />
<h1><center>July</center></h1><br />
<h3>Week 1</h3><br />
<p>Finnaly our constructs are already fixed. We ordered our synthetic gene for our main construct and ordered some parts from iGEM Headquarters. Our competent cell was also ready for transforming the other parts that we will use for our construct. We rehydrated BBa_K592025, BBa_J04450, BBa_C0040, BBa_B0017 and BBa_B0015, the colonies was growing well and ready for plasmid isolation. All of the plasmid was confirmed using restriction method and the result was positive. In this first week of July we learnt more about 3A assembly since we are ready to make our construct as well.</p><br />
<h3>Week 2</h3><br />
<p>We are ready to construct our building blocks. But first, we made some optimization method for double digest restriction reaction including for time incubation and reaction compotition. We also received our parts that we had ordered from iGEM headquarter BBa_K103006, BBa_K228004, BBa_K936024, BBa_K339010. We also tried to build our construct using two methods: standard assembly and 3A assembly. Still not work well. In this week we also elaborated and had brainstorming for our next method after our construct had been done.</p><br />
<h3>Week 3</h3><br />
<p>We were ready to try 3A assembly again, we made four different ligation composition to make sure which one will work well. After transformation, finally we got few colony in our petri disc, ready for plasmid isolation. After confirmation for all of the plasmid, we got no right colony yet. We then tried again using PSB1C3 with Rfp for the plasmid backbone instead of linearized plasmid to make it easier for colony selection using red-white screening. Still not got our right construct. Let the challenge begin and let’s do the best for it.</p><br />
<h3>Week 4</h3><br />
<p>Challenge accepted, we did our 3A again. We discussed about our failure for the 3A this two weeks and got some conclusion. Finally for this trial we got some colony which was the right construct BBa_K1387002. We then found the optimum ligation composition and reaction. Ready for another construct for reporter and converter module while waiting for synthetic gene for our main construct.</p><br />
<br><br />
<br />
<h1><center>August</center></h1><br />
<h3>Week 1</h3><br />
<p>Luckily, the primers were arrived. We ordered universal and spesific primer to check our construct using PCR technique. For optimization, we used gradient PCR technique. By using this technique, we were able to get a good results. Unfortunately, many of our construct were still in pSB1K3 backbone, so we must translocate it to pSB1C3 backbone.</p><br />
<h3>Week 2</h3><br />
<p align="justify">In this week we are trying to make our 3A assembly again since we still have some gene construct that must be finished before our synthetic gene comes. From the 3A assembly we got several colony that ready for plasmid isolation. After the confirmation through PCR methode, we did not get the right colony yet, so we must try the other compotition in ligation compotition, because we did not face any problem in transformation. After trying any ligation reaction , we got the optimal compotition for some construct, but the other still become our big mystery. In this week, we are also want to try the construct from UC Davis for Lc Cutinase BBa_K936020 and ethylene glycol converter module BBa_K936024. We did plasmid isolation from that part and transform it to the E.coli BL21 (DE3) We try to test the gene expression and test the product through SDS-PAGE method and we can see the protein band from both of the construct but not as the thick band. We also try to sequencing the positive construct in this week.</p><br />
<h3>Week 3</h3><br />
<p align="justify">Our target in this third week of august is to finish all of the construct except the construct that need synthetic gene due the parts submission deadline is coming. Restriction, purification gell (if needed), ligation, transformation, plasmid isolation, and confirmation is become our routine activities. In this week, we also learn more about how to express our protein, how to test our construct, how to measure the construct activities and did SDS-PAGE analysis.</p><br />
<h3>Week 4</h3><br />
<p align="justify">We focused to make a simple trial for our method to test the protein band of cutinase, and conversion methode, and discuss about our construct characterization especially for OmpA-LcCutinase construct (PET degradation module) as our main construct. We also discuss for synthetic gene cloning method. In this week we also focused on finishing our construct in PSB1C3 backbone and ready for shipment. Brainstoarming also become our routine activities because we found some failure in this week and feel not ready yet facing the parts submision deadline. We also try to elaborate how to make a modeling for our system and discuss with the expert.</p><br />
<br><br />
<br />
<h1><center>September</center></h1><br />
<h3>Week 1</h3><br />
<p align="justify">We have to prepare our parts submission form and check all of positive construct to make sure the condition of the construct is still good. In this week, synthetic gene, the thing that we waiting for one and half month finnaly arrived in our campuss. We try to make optimal reaction for our limited stock of synthetic gene. We try to clone our synthetic gene into some backbone including some of commercial clone vector and PSB1C3 of course. No collonies observed, so desperate and we discuss and evaluate our method and take a note of some mistake that we did in this method. Fortunatelly we have primer for our synthetic gene so we could do amplification using PCR for our synthetic gene. We also make new competent cell and trying electhrophoration transformation method instead of common heat shock methode. Still, no colonies observed.</p><br />
<h3>Week 2</h3><br />
<p align="justify">We try to clone our synthetic again. We must be succes in this week to step forward finishing our main construct. We try several method and try to use overnight ligation method. Finnaly, we got the positive colony, so happy. We also checked our plasmid again and found that some of our construct disappear but the other is still in good condition. We also make a optimization for PCR using touchdown and temperature gradient method to check the good temperature condition for the primer.</p><br />
<h3>Week 3</h3><br />
<p align="justify">We did PCR for construct confirmation for the collonies through crude PCR method using the optimal annealing temperature. We also did plasmid isolation for the positive colonies. We also make the method plan for PET degradation characterization using our construct and compared to UC Davis construct. We make some method for measurement including pH detection, ethylene glycol detection using dichromic acid method, and check the correlation between cell number in PET degradation rate.</p><br />
<h3>Week 4</h3><br />
<p align="justify">We submitted some of our parts including BBa_K1387000, BBa_K1387002 and BBa_K1387006 In the 11 days remaining for parts submission, we still try to build our construct and start to think the future development of our system. We also try to help Brawijaya team to cloning some of their parts and synthetic gene. We will start our parts characterization using several method including ethylene glycol assay using chromatic acid, pNPP assay to check the activity, growth curve, SEM, and qualitative assay using tributirin agar method.</p><br />
<br><br />
<br />
<h1><center>October</center></h1><br />
<h3>Week 1</h3><br />
<p align="justify">The sending parts week for second term. three parts is ready to send including BBa_K1387001, BBa_K1387006 and BBa_K1387005. Our dry lab team is also ready for modelling. We got some parameter to make the modelling. We also got some idea for our future system.</p><br />
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http://2014.igem.org/Team:ITB_Indonesia/nb-wetlab
Team:ITB Indonesia/nb-wetlab
2014-10-17T16:18:49Z
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<li>TEAM<br />
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<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
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<li>PROJECT<br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
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<li>NOTEBOOK<br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
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<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
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<br />
<br><br />
<h1><center>June</center></h1><br />
<h3>Week 1</h3><br />
<p>In the first week of June we observed the laboratory. We made a list of materials that we need for future wetlab activities. We prepared LB medium, searched the antibiotic optimization method and made competent cells using CCMB 80 buffer. We also sterilized all of the equipment needed. We already decided our idea that is engineer bacteria that can degrade plastic. Previously, we had four main ideas:<br><br />
1. Microalgae biofuel<br>2. Synthetic bacteria that can degrade plastic<br>3. Urine bio-sensor<br>4. Cyanide bio-sensor<br><br />
After competent cells were ready and antibiotic optimization had done then we had plan to try re-hydrated some parts and transform it to our <i> Escherichia coli </i> DH5α competent cells. We also had decided our construct so we would ready to order some synthetic parts and ordered it to iGEM HQ especially for some parts that not provided in the 2014 parts.</p><br />
<h3>Week 2</h3><br />
<p>This was our first time to see the iGEM distribution kit plate which was so exciting. We would make a trial to transform iGEM 2013 distribution kit using our competent cells. That transformation was not good on first, second, and third trial. Then we concluded that we must try the iGEM distribution kit plate 2014. We also tried to test our competent cells efficiency and we did not face any problem with our competent cells.</p><br />
<h3>Week 3</h3><br />
<p>Finally, we re-hydrated parts from iGEM kit plate 2014 which had many copies to make sure our method was optimal enough to transform those iGEM parts. We had already a plan to make four systems for our bacteria to degrade plastic. We divided our team into four groups. Each groups focused to each systems. We tried to transform BBa_K592025, BBa_B0015, BBa_B0034, BBa_I0040. The transformation did not have any validate colonies.</p><br />
<h3>Week 4</h3><br />
<p>We targeted to transform parts from iGEM distribution kit. Finally, we could see our red colony from RFP parts. So, we made liquid culture and wait until 16 hours. Then the medium turned into red. We tried to isolate the plasmid using Alkaline Lysis Method. The confirmation of the plasmids isolated were by electrophoresis, restriction and PCR method. The result was positive.</p><br />
<br><br />
<br />
<h1><center>July</center></h1><br />
<h3>Week 1</h3><br />
<p>Finnaly our constructs are already fixed. We ordered our synthetic gene for our main construct and ordered some parts from iGEM Headquarters. Our competent cell was also ready for transforming the other parts that we will use for our construct. We rehydrated BBa_K592025, BBa_J04450, BBa_C0040, BBa_B0017 and BBa_B0015, the colonies was growing well and ready for plasmid isolation. All of the plasmid was confirmed using restriction method and the result was positive. In this first week of July we learnt more about 3A assembly since we are ready to make our construct as well.</p><br />
<h3>Week 2</h3><br />
<p>We are ready to construct our building blocks. But first, we made some optimization method for double digest restriction reaction including for time incubation and reaction compotition. We also received our parts that we had ordered from iGEM headquarter BBa_K103006, BBa_K228004, BBa_K936024, BBa_K339010. We also tried to build our construct using two methods: standard assembly and 3A assembly. Still not work well. In this week we also elaborated and had brainstorming for our next method after our construct had been done.</p><br />
<h3>Week 3</h3><br />
<p>We were ready to try 3A assembly again, we made four different ligation composition to make sure which one will work well. After transformation, finally we got few colony in our petri disc, ready for plasmid isolation. After confirmation for all of the plasmid, we got no right colony yet. We then tried again using PSB1C3 with Rfp for the plasmid backbone instead of linearized plasmid to make it easier for colony selection using red-white screening. Still not got our right construct. Let the challenge begin and let’s do the best for it.</p><br />
<h3>Week 4</h3><br />
<p>Challenge accepted, we did our 3A again. We discussed about our failure for the 3A this two weeks and got some conclusion. Finally for this trial we got some colony which was the right construct BBa_K1387002. We then found the optimum ligation composition and reaction. Ready for another construct for reporter and converter module while waiting for synthetic gene for our main construct.</p><br />
<br><br />
<br />
<h1><center>August</center></h1><br />
<h3>Week 1</h3><br />
<p>Luckily, the primers were arrived. We ordered universal and spesific primer to check our construct using PCR technique. For optimization, we used gradient PCR technique. By using this technique, we were able to get a good results. Unfortunately, many of our construct were still in pSB1K3 backbone, so we must translocate it to pSB1C3 backbone.</p><br />
<h3>Week 2</h3><br />
<p>In this week we are trying to make our 3A assembly again since we still have some gene construct that must be finished before our synthetic gene comes. From the 3A assembly we got several colony that ready for plasmid isolation. After the confirmation through PCR methode, we did not get the right colony yet, so we must try the other compotition in ligation compotition, because we did not face any problem in transformation. After trying any ligation reaction , we got the optimal compotition for some construct, but the other still become our big mystery. In this week, we are also want to try the construct from UC Davis for Lc Cutinase BBa_K936020 and ethylene glycol converter module BBa_K936024. We did plasmid isolation from that part and transform it to the E.coli BL21 (DE3) We try to test the gene expression and test the product through SDS-PAGE method and we can see the protein band from both of the construct but not as the thick band. We also try to sequencing the positive construct in this week.</p><br />
<h3>Week 3</h3><br />
<p>Our target in this third week of august is to finish all of the construct except the construct that need synthetic gene due the parts submission deadline is coming. Restriction, purification gell (if needed), ligation, transformation, plasmid isolation, and confirmation is become our routine activities. In this week, we also learn more about how to express our protein, how to test our construct, how to measure the construct activities and did SDS-PAGE analysis.</p><br />
<h3>Week 4</h3><br />
<p>We focused to make a simple trial for our method to test the protein band of cutinase, and conversion methode, and discuss about our construct characterization especially for OmpA-LcCutinase construct (PET degradation module) as our main construct. We also discuss for synthetic gene cloning method. In this week we also focused on finishing our construct in PSB1C3 backbone and ready for shipment. Brainstoarming also become our routine activities because we found some failure in this week and feel not ready yet facing the parts submision deadline. We also try to elaborate how to make a modeling for our system and discuss with the expert.</p><br />
<br><br />
<br />
<h1><center>September</center></h1><br />
<h3>Week 1</h3><br />
<p>We have to prepare our parts submission form and check all of positive construct to make sure the condition of the construct is still good. In this week, synthetic gene, the thing that we waiting for one and half month finnaly arrived in our campuss. We try to make optimal reaction for our limited stock of synthetic gene. We try to clone our synthetic gene into some backbone including some of commercial clone vector and PSB1C3 of course. No collonies observed, so desperate and we discuss and evaluate our method and take a note of some mistake that we did in this method. Fortunatelly we have primer for our synthetic gene so we could do amplification using PCR for our synthetic gene. We also make new competent cell and trying electhrophoration transformation method instead of common heat shock methode. Still, no colonies observed.</p><br />
<h3>Week 2</h3><br />
<p>We try to clone our synthetic again. We must be succes in this week to step forward finishing our main construct. We try several method and try to use overnight ligation method. Finnaly, we got the positive colony, so happy. We also checked our plasmid again and found that some of our construct disappear but the other is still in good condition. We also make a optimization for PCR using touchdown and temperature gradient method to check the good temperature condition for the primer.</p><br />
<h3>Week 3</h3><br />
<p>We did PCR for construct confirmation for the collonies through crude PCR method using the optimal annealing temperature. We also did plasmid isolation for the positive colonies. We also make the method plan for PET degradation characterization using our construct and compared to UC Davis construct. We make some method for measurement including pH detection, ethylene glycol detection using dichromic acid method, and check the correlation between cell number in PET degradation rate.</p><br />
<h3>Week 4</h3><br />
<p>We submitted some of our parts including BBa_K1387000, BBa_K1387002 and BBa_K1387006 In the 11 days remaining for parts submission, we still try to build our construct and start to think the future development of our system. We also try to help Brawijaya team to cloning some of their parts and synthetic gene. We will start our parts characterization using several method including ethylene glycol assay using chromatic acid, pNPP assay to check the activity, growth curve, SEM, and qualitative assay using tributirin agar method.</p><br />
<br><br />
<br />
<h1><center>October</center></h1><br />
<h3>Week 1</h3><br />
<p>The sending parts week for second term. three parts is ready to send including BBa_K1387001, BBa_K1387006 and BBa_K1387005. Our dry lab team is also ready for modelling. We got some parameter to make the modelling. We also got some idea for our future system.</p><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Data
Team:ITB Indonesia/Data
2014-10-17T16:08:07Z
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<h1>A. Scaning Electron Microscope (SEM) Result Analysis</h1><br />
<p align="justify">For biodegrading experiments, we used 3x5 cm <sup> 2 </sup> bottle plastics with average weight 0.05 g. Then, we washed it several times with water and ethanol.</p><br />
<p align="justify">Plastic samples were applied just before we inoculated the bacteria on the medium. Bacterial culture was grown at 37<sup>0</sup>C in Luria-Broth medium. After 3 days of incubation, we washed the plastics with water and ethanol then dried for measurement of the weight loss.</p><br />
<p align="justify">Meanwhile, for UC Davis bacterial culture, we inoculated the bacteria in Luria-Broth medium supplemented with 170 ppm cloramphenicol. After an absorbance A600 nm of 0.6 was reached, we added 1% of arabinose and plastic samples. After 2 days of incubation, we washed the plastics with water and ethanol then dried for measurement of the weight loss.</p><br />
<p align="justify">We used medium supplemented with antibiotics and plastics sample as control on this experiment. The scaning electron microsccopy analysis of fractured surface of PET film was carried out using Scaning Electron Microscope. The surface of the treated PET samples were coated with conductive heavy metals such as gold/palladium.</p><br />
<palign="justify">SEM image shows that cracks were observed at the surface of a plastic samples (PET) after incubation of the bacterial culture. However, there was no significant weight decreased of the plastics samples. From this SEM study we concluded that both our LC Cut UC Davis and OmpA-LC-Cut Fussion ITB 2014 able to degrade PET plastics samples. Figure 1. shown the control sampel that we could not observe any cracks. Figure 2 and 3 shown the PET degradation by Lc cutinase from UC davis and from ITB_Indonesia.</p><br />
<br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/e/e9/Sem1.JPG"><br />
<p><small>Figure 1. Control sampel (PET plastic incubated in medium)</small></p><br />
</div><br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/9/92/Sem2.JPG"><br />
<p><small>Figure 2. PET plastic after treatment with ompA-LC-cutinase from ITB_Indonesia construct (BBa_K1387006)</small></p><br />
</div><br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/6/62/Sem3.JPG"><br />
<p><small>Figure 3. PET plastic after treatment with LC-cutinase UC Davis 2012 gene construct (Bba_K936020)</small></p><br />
</div><br />
<br />
<h3>References</h3><br />
<ol style="text-decoration:none;"><br />
<li>Sepperumal, Umaheswari, Murali Markandan, and Anbusaravanan Natarajan. 2013. electron microscopic studies of Polyethylene terepthalate degradation potential of Pseudomonas species. J. Microbiol. Biotech. Res. (1): 104-110</li><br />
<li>Mittal,Alok, R.K. Soni, Khrisna Dutt, and Swati Singh. 2010. Scaning electron microscopy study of hazardous waste flakes of polyethylene terephthalate (PET) by aminolysis and ammonolysis. Journal of Hazardous Materials Volume 178, Issues 1-3 </li><br />
</ol><br />
<br />
<br><br />
<h1>B. Ethylene Glikol Chromic Acid Assay</h1><br />
<p align="justify">Poly(ethylene terephthalate) (PET) is a plastic that is a polymer of ethylene terephthalate (C10H8O4) units (Sarker et al., 2010). Polyethylene terephthalate (PET) can be degrade via limited enzymatic hydrolysis of the ester bond of the polymer backbone, one enzyme that have been assesed for this purpose is cutinase (Ribitsch et al., 2012). When polyethylene terephthalate degraded, it will produce terephthalic acid and ethylene glycol (Venkatachalam et al., 2012). Ethylene glycol is one of alcohol compound. Oxidation of alcohols by strong oxidants such as chromic acid (K2Cr2O7) in suphuric acid(H2SO4) is possible, but differs depending on the degree of alcohol. If a reaction has occurred using chromic acid in sulphuric acid, there is a color change from orange to green. In our project we use chromic acid to detect ethylene glycol as product of the PET degradation by LC cutinase enzyme.</p><br />
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<div style="text-align: center;"><br />
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<p align="justify">Each sample used for measure the absorbance contain a fixed amount of chromic acid. The absorbance of the chromic acid will represent the total amount of ethylene glycol inversely. It means that if the absorbance is high, the amount of ethylene glycol in solution is low, and if the absorbance is low, the amount of ethylene glycol in solution is high. If the absorbance is high, the amount of chromic acid is high, which means that the amount of ethylene glycol being oxidized by chromic acid is low and vice versa. Both of LC Cutinase UC Davis and OmpA-LC Cutinase ITB_Indonesia shown the degrading activity of PET become ethylene glycol as once of the product.The optimum time to produce ethylene glycol is on the fourth hour after inoculation.</p><br />
<br />
<h3>References</h3><br />
<ol style="text-decoration:none;"><br />
<li>Ribitsch D, Enrique HA, Katrin G, Anita D, Sabine Z, Annemarie M, Rosario DR, Georg S, Karl G, Helmut S and Georg MG. 2012. A New Esterase from Thermobifida halotolerans Hydrolyses Polyethylene Terephthalate (PET) and Polylactic Acid (PLA). <i>Polymers. 4: 617-629</i></li><br />
<li>Sarker M, Aminul K, Mohammad MR, Mohammed M and ASMD Mohammad. 2011. Waste Polyethylene Terephthalate (PETE-1) Conversion into Liquid Fuel. <i>Journal of Fundamentals of Renewable Energy and Applications. 1</i></li><br />
<li>Venkatachalam S, Shilpa GN, Jayprakash VL, Prashant RG, Krishna R and Anil KK. 2012. Degradation and Recyclability of Poly (Ethylene Terephthalate). <i>Intech</i></li><br />
</ol><br />
<br />
<br><br />
<h1>C. pNPP assay</h1><br />
<p align="justify">Blastp program shows that LC cutinase shows high amino acid sequence similarity (54 to 60%) to lipase (Sulaiman, 2012). So, we tried to measure LC-cutinase activity using p-nitrophenylpalmitate (pNPP) as a substrate. pNPP typically used for measuring lipase activity.</p><br />
<p>In this assay, we used bacterial culture because in real application we would like to use bacterial culture to degrade PET directly. As negative control, we used <i>''E.coli'' </i> BL21(DE3) without plasmid. We also included LC-cutinase UC Davis 2012 gene construct (Bba_K936020) in this experiment. LC-cutinase UC Davis 2012 bacterial culture is induced after 2 hours of growth (after OD 600 nm ±0.6 was reached) using 1% of arabinose. All of the bacterial culture were harvested after 4 hours of growth.From ethylene glycol assay using chromic acid, we found that ompA-LC-cutinase from ITB_Indonesia construct (BBa_K1387006) has the highest activity after 4 hours of growth.</p><br />
<p align="justify">At first, we measured the activity at 60 degrees Celsius. We found that both ompA-LC-cutinase from ITB_Indonesia and LC-cutinase UC Davis 2012 still performed small activity. Then, we tried measure the activity at 25 degrees Celsius. The ompA-LC-cutinase from ITB_Indonesia’s activity was 0.001 U/mL. While, LC-cutinase UC Davis 2012 has ten fold higher (0.01 U/mL). The LC cutinase activity was determined based on the standard curve of p-nitrophenol. One unit of cutinase activity was defined as the amount of enzyme releasing 1 μmol pNP per minute under the assay conditions.</p><br />
<p align="justify">From this experiment, we are able to confirm that ompA-LC-cutinase from ITB_Indonesia successfully perform activity. But, its suggest that we still have to check the possible activity in different time of bacterial growth, expression rate, LC-cutinase presentation by ompA and activity assay using different substrate.</p><br />
<br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/9/90/Sem6.JPG"><br />
<p><small>Figure 1. LC Cutinase activity. The enzymatic activity was determined at 60C in assay condition using pNP-palmitate (C16) as a substrate. The experiment was carried out twice.</small></p><br />
</div><br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/a/a1/Sem7.JPG"><br />
<p><small>Figure 2. LC Cutinase activity. The enzymatic activity was determined at 25 C in assay condition using pNP-palmitate (C16) as a substrate. The experiment was carried out twice.</small></p><br />
</div><br />
<p align"justify">Small activity of LC-Cutinase maybe due to their oligomeric state. Tethering protein to cell surface may affect folding of protein and also their biological function (Park,2010). These suggest us to look futher detail on folding and displaying state of LC cutinase.</p><br />
<br />
<p align="justify">There are several things that should be noted when we designate fusion strategy of passanger protein or protein of interest with the outer membrane of protein. Linking the different passanger proteins to the same outer membrane protein may result in different translocation. The characteristic of passanger protein affects the translocation. Its characteristic such as formation of disulfide bridge, total residue of hydrophobic aminoacid etc. </p><br />
<p align="justify">So there are four requirements have to be met for constructing well designed surface display. First is, it has to possesses efficient signal peptide. Second, it should have strong anchor to keep the fusion protein from detachment. Third, it should be compatible to the sequence that is inserted or fused. The last is it should resisstant to the attack of the protease from medium or periplasmic membrane. </p><br />
<br />
<h3>Reference</h3><br />
<ol style="text-decoration:none;"><br />
<li>Sulaiman, Sintawee, SayaYamat, EikoKanaya, Joong-Jae Kim, Yuichi Koga, Kazufumi Takano and Shigenori Kanaya. 2012. Isolation of a Novel Cutinase Homolog with Polyethylene Terephthalate-Degrading Activity from Leaf-Branch Compost by Using a Metagenomic Approach. <i>Applied and Environmental Microbiology p 1556-1562.</i></li><br />
<li>Lee, Sang Yup., Choi, Jong Hyun., Xu, Zhaohui. 2003. “Microbial Cell-Surface Display” <i>Elsevier</i></li><br />
</ol><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
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<br><br />
<h1>A. Scaning Electron Microscope (SEM) Result Analysis</h1><br />
<p align="justify">For biodegrading experiments, we used 3x5 cm <sup> 2 </sup> bottle plastics with average weight 0.05 g. Then, we washed it several times with water and ethanol.</p><br />
<p align="justify">Plastic samples were applied just before we inoculated the bacteria on the medium. Bacterial culture was grown at 37<sup>0</sup>C in Luria-Broth medium. After 3 days of incubation, we washed the plastics with water and ethanol then dried for measurement of the weight loss.</p><br />
<p align="justify">Meanwhile, for UC Davis bacterial culture, we inoculated the bacteria in Luria-Broth medium supplemented with 170 ppm cloramphenicol. After an absorbance A600 nm of 0.6 was reached, we added 1% of arabinose and plastic samples. After 2 days of incubation, we washed the plastics with water and ethanol then dried for measurement of the weight loss.</p><br />
<p align="justify">We used medium supplemented with antibiotics and plastics sample as control on this experiment. The scaning electron microsccopy analysis of fractured surface of PET film was carried out using Scaning Electron Microscope. The surface of the treated PET samples were coated with conductive heavy metals such as gold/palladium.</p><br />
<palign="justify">SEM image shows that cracks were observed at the surface of a plastic samples (PET) after incubation of the bacterial culture. However, there was no significant weight decreased of the plastics samples. From this SEM study we concluded that both our LC Cut UC Davis and OmpA-LC-Cut Fussion ITB 2014 able to degrade PET plastics samples. Figure 1. shown the control sampel that we could not observe any cracks. Figure 2 and 3 shown the PET degradation by Lc cutinase from UC davis and from ITB_Indonesia.</p><br />
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<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/e/e9/Sem1.JPG"><br />
<p><small>Figure 1. Control sampel (PET plastic incubated in medium)</small></p><br />
</div><br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/9/92/Sem2.JPG"><br />
<p><small>Figure 2. PET plastic after treatment with ompA-LC-cutinase from ITB_Indonesia construct (BBa_K1387006)</small></p><br />
</div><br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/6/62/Sem3.JPG"><br />
<p><small>Figure 3. PET plastic after treatment with LC-cutinase UC Davis 2012 gene construct (Bba_K936020)</small></p><br />
</div><br />
<br />
<h3>References</h3><br />
<ol style="text-decoration:none;"><br />
<li>Sepperumal, Umaheswari, Murali Markandan, and Anbusaravanan Natarajan. 2013. electron microscopic studies of Polyethylene terepthalate degradation potential of Pseudomonas species. J. Microbiol. Biotech. Res. (1): 104-110</li><br />
<li>Mittal,Alok, R.K. Soni, Khrisna Dutt, and Swati Singh. 2010. Scaning electron microscopy study of hazardous waste flakes of polyethylene terephthalate (PET) by aminolysis and ammonolysis. Journal of Hazardous Materials Volume 178, Issues 1-3 </li><br />
</ol><br />
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<br><br />
<h1>B. Ethylene Glikol Chromic Acid Assay</h1><br />
<p align="justify">Poly(ethylene terephthalate) (PET) is a plastic that is a polymer of ethylene terephthalate (C10H8O4) units (Sarker et al., 2010). Polyethylene terephthalate (PET) can be degrade via limited enzymatic hydrolysis of the ester bond of the polymer backbone, one enzyme that have been assesed for this purpose is cutinase (Ribitsch et al., 2012). When polyethylene terephthalate degraded, it will produce terephthalic acid and ethylene glycol (Venkatachalam et al., 2012). Ethylene glycol is one of alcohol compound. Oxidation of alcohols by strong oxidants such as chromic acid (K2Cr2O7) in suphuric acid(H2SO4) is possible, but differs depending on the degree of alcohol. If a reaction has occurred using chromic acid in sulphuric acid, there is a color change from orange to green. In our project we use chromic acid to detect ethylene glycol as product of the PET degradation by LC cutinase enzyme.</p><br />
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<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/2/2e/Sem5.JPG"><br />
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<img src="https://static.igem.org/mediawiki/2014/2/23/Sem4.JPG"><br />
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<p align="justify">Each sample used for measure the absorbance contain a fixed amount of chromic acid. The absorbance of the chromic acid will represent the total amount of ethylene glycol inversely. It means that if the absorbance is high, the amount of ethylene glycol in solution is low, and if the absorbance is low, the amount of ethylene glycol in solution is high. If the absorbance is high, the amount of chromic acid is high, which means that the amount of ethylene glycol being oxidized by chromic acid is low and vice versa. Both of LC Cutinase UC Davis and OmpA-LC Cutinase ITB_Indonesia shown the degrading activity of PET become ethylene glycol as once of the product.The optimum time to produce ethylene glycol is on the fourth hour after inoculation.</p><br />
<br />
<h3>References</h3><br />
<ol style="text-decoration:none;"><br />
<li>Ribitsch D, Enrique HA, Katrin G, Anita D, Sabine Z, Annemarie M, Rosario DR, Georg S, Karl G, Helmut S and Georg MG. 2012. A New Esterase from Thermobifida halotolerans Hydrolyses Polyethylene Terephthalate (PET) and Polylactic Acid (PLA). <i>Polymers. 4: 617-629</i></li><br />
<li>Sarker M, Aminul K, Mohammad MR, Mohammed M and ASMD Mohammad. 2011. Waste Polyethylene Terephthalate (PETE-1) Conversion into Liquid Fuel. <i>Journal of Fundamentals of Renewable Energy and Applications. 1</i></li><br />
<li>Venkatachalam S, Shilpa GN, Jayprakash VL, Prashant RG, Krishna R and Anil KK. 2012. Degradation and Recyclability of Poly (Ethylene Terephthalate). <i>Intech</i></li><br />
</ol><br />
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<br><br />
<h1>C. pNPP assay</h1><br />
<p align="justify">Blastp program shows that LC cutinase shows high amino acid sequence similarity (54 to 60%) to lipase (Sulaiman, 2012). So, we tried to measure LC-cutinase activity using p-nitrophenylpalmitate (pNPP) as a substrate. pNPP typically used for measuring lipase activity.</p><br />
<p>In this assay, we used bacterial culture because in real application we would like to use bacterial culture to degrade PET directly. As negative control, we used <i>''E.coli'' </i> BL21(DE3) without plasmid. We also included LC-cutinase UC Davis 2012 gene construct (Bba_K936020) in this experiment. LC-cutinase UC Davis 2012 bacterial culture is induced after 2 hours of growth (after OD 600 nm ±0.6 was reached) using 1% of arabinose. All of the bacterial culture were harvested after 4 hours of growth.From ethylene glycol assay using chromic acid, we found that ompA-LC-cutinase from ITB_Indonesia construct (BBa_K1387006) has the highest activity after 4 hours of growth.</p><br />
<p align="justify">At first, we measured the activity at 60 degrees Celsius. We found that both ompA-LC-cutinase from ITB_Indonesia and LC-cutinase UC Davis 2012 still performed small activity. Then, we tried measure the activity at 25 degrees Celsius. The ompA-LC-cutinase from ITB_Indonesia’s activity was 0.001 U/mL. While, LC-cutinase UC Davis 2012 has ten fold higher (0.01 U/mL). The LC cutinase activity was determined based on the standard curve of p-nitrophenol. One unit of cutinase activity was defined as the amount of enzyme releasing 1 μmol pNP per minute under the assay conditions.</p><br />
<p align="justify">From this experiment, we are able to confirm that ompA-LC-cutinase from ITB_Indonesia successfully perform activity. But, its suggest that we still have to check the possible activity in different time of bacterial growth, expression rate, LC-cutinase presentation by ompA and activity assay using different substrate.</p><br />
<br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/9/90/Sem6.JPG"><br />
<p><small>Figure 1. LC Cutinase activity. The enzymatic activity was determined at 60C in assay condition using pNP-palmitate (C16) as a substrate. The experiment was carried out twice.</small></p><br />
</div><br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/a/a1/Sem7.JPG"><br />
<p><small>Figure 2. LC Cutinase activity. The enzymatic activity was determined at 25 C in assay condition using pNP-palmitate (C16) as a substrate. The experiment was carried out twice.</small></p><br />
</div><br />
<p align"justify">Small activity of LC-Cutinase maybe due to their oligomeric state. Tethering protein to cell surface may affect folding of protein and also their biological function (Park,2010). These suggest us to look futher detail on folding and displaying state of LC cutinase.</p><br />
<br />
<p align="justify">There are several things that should be noted when we designate fusion strategy of passanger protein or protein of interest with the outer membrane of protein. Linking the different passanger proteins to the same outer membrane protein may result in different translocation. The characteristic of passanger protein affects the translocation. Its characteristic such as formation of disulfide bridge, total residue of hydrophobic aminoacid etc. </p><br />
<p align="justify">So there are four requirements have to be met for constructing well designed surface display. First is, it has to possesses efficient signal peptide, Second, it should have strong anchor to keep the fusion protein from detachment. Third, it should be compatible to the sequence that is inserted or fused. The last is it should resisstant to the attack of the protease from medium or periplasmic membrane. </p><br />
<br />
<h3>Reference</h3><br />
<ol style="text-decoration:none;"><br />
<li>Sulaiman, Sintawee, SayaYamat, EikoKanaya, Joong-Jae Kim, Yuichi Koga, Kazufumi Takano and Shigenori Kanaya. 2012. Isolation of a Novel Cutinase Homolog with Polyethylene Terephthalate-Degrading Activity from Leaf-Branch Compost by Using a Metagenomic Approach. <i>Applied and Environmental Microbiology p 1556-1562.</i></li><br />
<li>Lee, Sang Yup., Choi, Jong Hyun., Xu, Zhaohui. 2003. “Microbial Cell-Surface Display” <i>Elsevier</i></li><br />
</ol><br />
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http://2014.igem.org/Team:ITB_Indonesia/nb-wetlab
Team:ITB Indonesia/nb-wetlab
2014-10-17T15:56:55Z
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<h1><center>June</center></h1><br />
<h3>Week 1</h3><br />
<p>In the first week of June we observed the laboratory. We made a list of materials that we need for future wetlab activities. We prepared LB medium, searched the antibiotic optimization method and made competent cells using CCMB 80 buffer. We also sterilized all of the equipment needed. We already decided our idea that is engineer bacteria that can degrade plastic. Previously, we had four main ideas:<br><br />
1. Microalgae biofuel<br>2. Synthetic bacteria that can degrade plastic<br>3. Urine bio-sensor<br>4. Cyanide bio-sensor<br><br />
After competent cells were ready and antibiotic optimization had done then we had plan to try re-hydrated some parts and transform it to our ''Escherichia coli'' DH5α competent cells. We also had decided our construct so we would ready to order some synthetic parts and ordered it to iGEM HQ especially for some parts that not provided in the 2014 parts.</p><br />
<h3>Week 2</h3><br />
<p>This was our first time to see the iGEM distribution kit plate which was so exciting. We would make a trial to transform iGEM 2013 distribution kit using our competent cells. That transformation was not good on first, second, and third trial. Then we concluded that we must try the iGEM distribution kit plate 2014. We also tried to test our competent cells efficiency and we did not face any problem with our competent cells.</p><br />
<h3>Week 3</h3><br />
<p>Finally, we re-hydrated parts from iGEM kit plate 2014 which had many copies to make sure our method was optimal enough to transform those iGEM parts. We had already a plan to make four systems for our bacteria to degrade plastic. We divided our team into four groups. Each groups focused to each systems. We tried to transform BBa_K592025, BBa_B0015, BBa_B0034, BBa_I0040. The transformation did not have any validate colonies.</p><br />
<h3>Week 4</h3><br />
<p>We targeted to transform parts from iGEM distribution kit. Finally, we could see our red colony from RFP parts. So, we made liquid culture and wait until 16 hours. Then the medium turned into red. We tried to isolate the plasmid using Alkaline Lysis Method. The confirmation of the plasmids isolated were by electrophoresis, restriction and PCR method. The result was positive.</p><br />
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<h1><center>July</center></h1><br />
<h3>Week 1</h3><br />
<p>Finnaly our constructs are already fixed. We ordered our synthetic gene for our main construct and ordered some parts from iGEM Headquarters. Our competent cell was also ready for transforming the other parts that we will use for our construct. We rehydrated BBa_K592025, BBa_J04450, BBa_C0040, BBa_B0017 and BBa_B0015, the colonies was growing well and ready for plasmid isolation. All of the plasmid was confirmed using restriction method and the result was positive. In this first week of July we learnt more about 3A assembly since we are ready to make our construct as well.</p><br />
<h3>Week 2</h3><br />
<p>We are ready to construct our building blocks. But first, we made some optimization method for double digest restriction reaction including for time incubation and reaction compotition. We also received our parts that we had ordered from iGEM headquarter BBa_K103006, BBa_K228004, BBa_K936024, BBa_K339010. We also tried to build our construct using two methods: standard assembly and 3A assembly. Still not work well. In this week we also elaborated and had brainstorming for our next method after our construct had been done.</p><br />
<h3>Week 3</h3><br />
<p>We were ready to try 3A assembly again, we made four different ligation composition to make sure which one will work well. After transformation, finally we got few colony in our petri disc, ready for plasmid isolation. After confirmation for all of the plasmid, we got no right colony yet. We then tried again using PSB1C3 with Rfp for the plasmid backbone instead of linearized plasmid to make it easier for colony selection using red-white screening. Still not got our right construct. Let the challenge begin and let’s do the best for it.</p><br />
<h3>Week 4</h3><br />
<p>Challenge accepted, we did our 3A again. We discussed about our failure for the 3A this two weeks and got some conclusion. Finally for this trial we got some colony which was the right construct BBa_K1387002. We then found the optimum ligation composition and reaction. Ready for another construct for reporter and converter module while waiting for synthetic gene for our main construct.</p><br />
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<h1><center>August</center></h1><br />
<h3>Week 1</h3><br />
<p>Luckily, the primers were arrived. We ordered universal and spesific primer to check our construct using PCR technique. For optimization, we used gradient PCR technique. By using this technique, we were able to get a good results. Unfortunately, many of our construct were still in pSB1K3 backbone, so we must translocate it to pSB1C3 backbone.</p><br />
<h3>Week 2</h3><br />
<p>In this week we are trying to make our 3A assembly again since we still have some gene construct that must be finished before our synthetic gene comes. From the 3A assembly we got several colony that ready for plasmid isolation. After the confirmation through PCR methode, we did not get the right colony yet, so we must try the other compotition in ligation compotition, because we did not face any problem in transformation. After trying any ligation reaction , we got the optimal compotition for some construct, but the other still become our big mystery. In this week, we are also want to try the construct from UC Davis for Lc Cutinase BBa_K936020 and ethylene glycol converter module BBa_K936024. We did plasmid isolation from that part and transform it to the E.coli BL21 (DE3) We try to test the gene expression and test the product through SDS-PAGE method and we can see the protein band from both of the construct but not as the thick band. We also try to sequencing the positive construct in this week.</p><br />
<h3>Week 3</h3><br />
<p>Our target in this third week of august is to finish all of the construct except the construct that need synthetic gene due the parts submission deadline is coming. Restriction, purification gell (if needed), ligation, transformation, plasmid isolation, and confirmation is become our routine activities. In this week, we also learn more about how to express our protein, how to test our construct, how to measure the construct activities and did SDS-PAGE analysis.</p><br />
<h3>Week 4</h3><br />
<p>We focused to make a simple trial for our method to test the protein band of cutinase, and conversion methode, and discuss about our construct characterization especially for OmpA-LcCutinase construct (PET degradation module) as our main construct. We also discuss for synthetic gene cloning method. In this week we also focused on finishing our construct in PSB1C3 backbone and ready for shipment. Brainstoarming also become our routine activities because we found some failure in this week and feel not ready yet facing the parts submision deadline. We also try to elaborate how to make a modeling for our system and discuss with the expert.</p><br />
<br><br />
<br />
<h1><center>September</center></h1><br />
<h3>Week 1</h3><br />
<p>We have to prepare our parts submission form and check all of positive construct to make sure the condition of the construct is still good. In this week, synthetic gene, the thing that we waiting for one and half month finnaly arrived in our campuss. We try to make optimal reaction for our limited stock of synthetic gene. We try to clone our synthetic gene into some backbone including some of commercial clone vector and PSB1C3 of course. No collonies observed, so desperate and we discuss and evaluate our method and take a note of some mistake that we did in this method. Fortunatelly we have primer for our synthetic gene so we could do amplification using PCR for our synthetic gene. We also make new competent cell and trying electhrophoration transformation method instead of common heat shock methode. Still, no colonies observed.</p><br />
<h3>Week 2</h3><br />
<p>We try to clone our synthetic again. We must be succes in this week to step forward finishing our main construct. We try several method and try to use overnight ligation method. Finnaly, we got the positive colony, so happy. We also checked our plasmid again and found that some of our construct disappear but the other is still in good condition. We also make a optimization for PCR using touchdown and temperature gradient method to check the good temperature condition for the primer.</p><br />
<h3>Week 3</h3><br />
<p>We did PCR for construct confirmation for the collonies through crude PCR method using the optimal annealing temperature. We also did plasmid isolation for the positive colonies. We also make the method plan for PET degradation characterization using our construct and compared to UC Davis construct. We make some method for measurement including pH detection, ethylene glycol detection using dichromic acid method, and check the correlation between cell number in PET degradation rate.</p><br />
<h3>Week 4</h3><br />
<p>We submitted some of our parts including BBa_K1387000, BBa_K1387002 and BBa_K1387006 In the 11 days remaining for parts submission, we still try to build our construct and start to think the future development of our system. We also try to help Brawijaya team to cloning some of their parts and synthetic gene. We will start our parts characterization using several method including ethylene glycol assay using chromatic acid, pNPP assay to check the activity, growth curve, SEM, and qualitative assay using tributirin agar method.</p><br />
<br><br />
<br />
<h1><center>October</center></h1><br />
<h3>Week 1</h3><br />
<p>The sending parts week for second term. three parts is ready to send including BBa_K1387001, BBa_K1387006 and BBa_K1387005. Our dry lab team is also ready for modelling. We got some parameter to make the modelling. We also got some idea for our future system.</p><br />
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http://2014.igem.org/Team:ITB_Indonesia/Achievement
Team:ITB Indonesia/Achievement
2014-10-17T14:37:34Z
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<br><br />
<h1>Achievements</h1><br />
<ol><br />
<li><b>Designed several new BioBricks</b></li><br />
<li><b>Characterised our BioBricks <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K1387006">BBa_K1387006</a></b></li><br />
<li><b>Helped UB_INDONESIA 2014 Team by debugging their construct (<a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K1367006">BBa_K1367006</a> and <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K1367007">BBa_K1367007</a>)</b></li><br />
<li><b>Characterised an existing BioBrick Device <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K936020">(BBa_K936020)</a></b></li><br />
<li><b>Developed new method using chromic acid for ethylene glycol assay</b></li><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Achievement
Team:ITB Indonesia/Achievement
2014-10-17T14:26:56Z
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<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Achievements</h1><br />
<ol><br />
<li><b>Designed several new BioBricks</b></li><br />
<li><b>Characterised our BioBricks <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K1387006">BBa_K1387006</a></b></li><br />
<li><b>Helped UB_INDONESIA 2014 Team by debugging their construct (BBa_K1367006 and BBa_K1367007)</b></li><br />
<li><b>Characterised an existing BioBrick Device (BBa_K936020)</b></li><br />
<li><b>Developed new method using chromic acid for ethylene glycol assay</b></li><br />
</ol><br />
<br><br />
<br><br />
<br />
<div id="sponsor"></div><br />
</div><br />
</div><br />
</body><br />
</html></div>
Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Team
Team:ITB Indonesia/Team
2014-10-17T13:36:20Z
<p>Erawijantari: </p>
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<div id="putih"> <div id="logo-igem" onclick="window.location.href='https://2014.igem.org/'"></div><br />
<div id="center"><br />
<div id="logo"></div><br />
<br />
<div id="header"></div><br />
<br />
<div id="blockNav"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<div class="contenner"><br />
<br />
<h1>Team Leader</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/0/06/Team_Leader_ITB.JPG" width="270px" height="333px" style="image-orientation: 90deg;"/></div><br />
<div class="desc"><p>The leader of ITB_Indonesia 2014. He keep all of members spirit. Dedicated and enthusiastic to genetic engineering and synthetic biology.</p><br />
<br><br><br><br><br><br><br><br><br><br><br />
<strong>Joko Pebrianto Trinugroho</strong><br>School of Life Science and Technology<br />
</div><br />
</div> <br />
<div class="contenner"><br />
<h1>Public Relation</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/0/0a/Public_Relation_Mardalisa.JPG" width="270px" height="333px" /></div><br />
<div class="desc"><p>Role of public relation in ITB_Indonesia 2014. Public Image strategy, outsearch events, media relations, social media, Handling emergencies.</p><br />
<br><br><br><br><br><br><br><br><br><br />
<strong>Mardalisa</strong><br>School of Life Science and Technology</div><br />
</div> <br />
<div class="contenner"> <br />
<h1>Creative</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/d/d2/Tim_Kreatif.jpg" width="270px" height="360px" /></div><br />
<div class="desc"><p>Most creative peoples in team</p><br />
<br><br><br><br><br><br><br><br><br><br><br />
<strong>Fania Feby Ramadhani</strong><br />
<br><strong>Elfa Norisda Aulianisa</strong><br>School of Life Science and Technology<br />
<br />
<br />
<br><br><strong>Muhammad Musyafa Syahbid</strong><br>School of Architecture, Planning, and Policy Development<br />
<br />
</div><br />
</div> <br />
<div class="contenner"><br />
<h1>Idea Team</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/9/9d/Tim_ide.JPG" width="270px" height="333px" /></div><br />
<div class="desc"><p>Idea Team acts as a mastermind in a construction of a novel device for project of team ITB_Indonesia 2014. Besides that, we also try to create a new improvement for the construction of our novel device and develop a new characterization technique. This improvement is hopefully will increase the efficiency and effectivity of the device and the new characterization technique is expected to minimize the budget of research, easily to carry out, and give more accurate result.</p><br />
<br><br><br><br />
<strong>Tirta Widi Gilang Citradi</strong><br><br />
<strong>Christian Heryakusuma</strong><br>School of Life Sciences and Technology<br><br><br />
<strong>Yovin</strong><br>Faculty of Mathematics and Natural Science<br><br />
</div><br />
</div> <br />
<div class="contenner"><br />
<h1>Sponsorship</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/2/29/Sponsorshipitb.JPG" width="270px" height="338px" /></div><br />
<div class="desc"><p>Acquisition of external fund to cover the project expenses, generate in-kind sponsorships, research corporation that would be interested in out project, prepare and submit excellent sponsorship prorposal. Establish and maintain good relationships with corporate representatives, take responsibility for the day to day fund raising, generate revenue through selling souvenir , food, laboratory lab coat, shirt and pen that can promote to socialized our project.</p><br />
<br><br><br><br />
<strong>Lance Rosa Karo-Karo</strong><br><br />
<strong>Oktira Roka Aji</strong><br><br />
<strong>Kenia Permata Sukma</strong><br><br />
<strong>Wiyudi Gomulya</strong><br><br />
<strong>I Dewa Gde Sathya Deva</strong><br>School of Life Sciences and Technology<br><br><br />
</div><br />
</div> <br />
<div class="contenner"><br />
<h1>Human Practice</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/9/9b/Human_Practice.jpg" width="270px" height="360px"/></div><br />
<div class="desc"><p>We are HUMAN PRACTISE! Theory without us is nothing. We introduce synthetic biology, make collaboration, make project documentation, make synthethic biology become enjoyable science, and spread happiness :)</p><br />
<br><br><br><br><br><br><br><br><br />
<strong>Nimas Ghasani</strong><br><br />
<strong>Risma Wiharyanti</strong><br><br />
<strong>Bakhtiar Hermawan</strong><br><br />
<strong>Wuddan Nadhirah Rodiana</strong><br>School of Life Sciences and Technology<br><br><br />
<strong>Elfina Marchantia</strong><br>Faculty of Mathematics and Natural Science<br><br><br />
</div><br />
</div> <br />
<div class="contenner"><br />
<h1>Wetlab</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/b/b7/WetLab.JPG" width="270px" height="333px"/></div><br />
<div class="desc"><p>They are “The Wetlabors” (wet lab – labor), some people say it because they always work hard to do the wet lab project. But they never hesitate to ask for help to the other members to do the Super Wet lab Project.</p><br />
<br><br><br><br><br><br><br><br><br><br><br><br><br />
<strong>Tri Ekawati Heryanto</strong><br><br />
<strong>Pande Putu Erawijantari</strong><br>School of Life Sciences and Technology<br><br><br />
</div><br />
</div> <br />
<div class="contenner"><br />
<h1>Wiki</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/9/9b/10708170_10202668676788565_1895796730_n.jpg" width="270px" height="360px" /></div><br />
<div class="desc"><p>Manage, Create and implement wiki's website</p><br />
<br><br><br><br><br><br><br><br><br><br><br><br><br />
<strong>Rizky Kusumah</strong><br><br />
<strong>Muhammad Hariomurti Mardikusumo</strong><br>School of Electrical Engineering and Informatics<br><br><br />
</div><br />
</div> <br />
<div class="contenner"> <br />
<br><br />
<h1>Team Advisor</h1><br />
<div class="image2"><img src="https://static.igem.org/mediawiki/igem.org/6/6a/2013_ITB_Indonesia-adv2.jpg" width="154px" height="205px" /></div><br />
<div class="desc2"><strong>Sony Suhandono, Ph.D</strong><p>Genetics and Molecular Biotechnology Research Group<br><br />
School of Life Sciences and Technology <br><br />
Institut Teknologi Bandung</p></div><br />
</div> <br><br />
<div><br />
<div class="image2"><img src="https://static.igem.org/mediawiki/igem.org/8/83/2013_ITB_Indonesia-adv1.jpg" width="154px" height="205px" /></div><br />
<div class="desc2"><strong>Maelita Ramdani Moeis, Ph.D</strong><p>Genetics and Molecular Biotechnology Research Group<br><br />
School of Life Sciences and Technology <br><br />
Institut Teknologi Bandung</p></div><br />
</div> <br><br />
<div><br />
<div class="image2"><img src="https://static.igem.org/mediawiki/2014/8/8a/Dr_Dessy_Natalia.jpg" width="154px" height="205px" /></div><br />
<div class="desc2"><strong>Dessy Natalia, Ph.D</strong><p>Biochemistry Research Division<br><br />
Faculty of Mathematics and Natural Sciences<br><br />
Institut Teknologi Bandung</p></div><br />
</div> <br><br />
<div><br />
<div class="image2"><img src="https://static.igem.org/mediawiki/2014/9/98/Dr_Mochamad_Apri.jpg" width="154px" height="205px" /></div><br />
<div class="desc2"><strong>Mochamad Apri, Ph.D</strong><p>Industrial and Financial Mathematics Research Division<br><br />
Faculty of Mathematics and Natural Sciences<br><br />
Institut Teknologi Bandung</p></div><br />
</div> <br><br />
<div><br />
<div class="image2"><img src="https://static.igem.org/mediawiki/2014/5/57/Dr_Indra_Wibowo.jpg" width="154px" height="205px" /></div><br />
<div class="desc2"><strong>Indra Wibowo, Ph.D</strong><p>Animal Physiology, Developmentand Biomedical Research Group<br><br />
School of Life Sciences and Technology <br><br />
Institut Teknologi Bandung</p></div><br />
</div> <br />
<h1>Instructor</h1><br />
<div class="image2"><img src="https://static.igem.org/mediawiki/parts/5/52/ITB-aridwijayanti.JPG" width="154px" height="205px" /></div><br />
<div class="desc2"><strong>Ari Dwijayanti, M.Si</strong><p>Genetics and Molecular Biotechnology Research Group<br><br />
School of Life Sciences and Technology <br><br />
Institut Teknologi Bandung</p></div><br />
</div> <br><br />
<br />
<br><br />
<br />
<div id="sponsor"></div><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Team
Team:ITB Indonesia/Team
2014-10-17T13:32:20Z
<p>Erawijantari: </p>
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<div id="putih"> <div id="logo-igem" onclick="window.location.href='https://2014.igem.org/'"></div><br />
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<br />
<div id="header"></div><br />
<br />
<div id="blockNav"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<div class="contenner"><br />
<br />
<h1>Team Leader</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/0/06/Team_Leader_ITB.JPG" width="270px" height="333px" style="image-orientation: 90deg;"/></div><br />
<div class="desc"><p>The leader of ITB_Indonesia 2014. He keep all of members spirit. Dedicated and enthusiastic to genetic engineering and synthetic biology.</p><br />
<br><br><br><br><br><br><br><br><br><br><br />
<strong>Joko Pebrianto Trinugroho</strong><br>School of Life Science and Technology<br />
</div><br />
</div> <br />
<div class="contenner"><br />
<h1>Public Relation</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/0/0a/Public_Relation_Mardalisa.JPG" width="270px" height="333px" /></div><br />
<div class="desc"><p>Role of public relation in ITB_Indonesia 2014. Public Image strategy, outsearch events, media relations, social media, Handling emergencies.</p><br />
<br><br><br><br><br><br><br><br><br><br />
<strong>Mardalisa</strong><br>School of Life Science and Technology</div><br />
</div> <br />
<div class="contenner"> <br />
<h1>Creative</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/d/d2/Tim_Kreatif.jpg" width="270px" height="360px" /></div><br />
<div class="desc"><p>Most creative peoples in team</p><br />
<br><br><br><br><br><br><br><br><br><br><br />
<strong>Fania Feby Ramadhani</strong><br />
<br><strong>Elfa Norisda Aulianisa</strong><br>School of Life Science and Technology<br />
<br />
<br />
<br><br><strong>Muhammad Musyafa Syahbid</strong><br>School of Architecture, Planning, and Policy Development<br />
<br />
</div><br />
</div> <br />
<div class="contenner"><br />
<h1>Idea Team</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/9/9d/Tim_ide.JPG" width="270px" height="333px" /></div><br />
<div class="desc"><p>Idea Team acts as a mastermind in a construction of a novel device for project of team ITB_Indonesia 2014. Besides that, we also try to create a new improvement for the construction of our novel device and develop a new characterization technique. This improvement is hopefully will increase the efficiency and effectivity of the device and the new characterization technique is expected to minimize the budget of research, easily to carry out, and give more accurate result.</p><br />
<br><br><br><br />
<strong>Tirta Widi Gilang Citradi</strong><br><br />
<strong>Christian Heryakusuma</strong><br>School of Life Sciences and Technology<br><br><br />
<strong>Yovin</strong><br>Faculty of Mathematics and Natural Science<br><br />
</div><br />
</div> <br />
<div class="contenner"><br />
<h1>Sponsorship</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/2/29/Sponsorshipitb.JPG" width="270px" height="338px" /></div><br />
<div class="desc"><p>Acquisition of external fund to cover the project expenses, generate in-kind sponsorships, research corporation that would be interested in out project, prepare and submit excellent sponsorship prorposal. Establish and maintain good relationships with corporate representatives, take responsibility for the day to day fund raising, generate revenue through selling souvenir , food, laboratory lab coat, shirt and pen that can promote to socialized our project.</p><br />
<br><br><br><br />
<strong>Lance Rosa Karo-Karo</strong><br><br />
<strong>Oktira Roka Aji</strong><br><br />
<strong>Kenia Permata Sukma</strong><br><br />
<strong>Wiyudi Gomulya</strong><br><br />
<strong>I Dewa Gde Sathya Deva</strong><br>School of Life Sciences and Technology<br><br><br />
</div><br />
</div> <br />
<div class="contenner"><br />
<h1>Human Practice</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/9/9b/Human_Practice.jpg" width="270px" height="360px"/></div><br />
<div class="desc"><p>We are HUMAN PRACTISE! Theory without us is nothing. We introduce synthetic biology, make collaboration, make project documentation, make synthethic biology become enjoyable science, and spread happiness :)</p><br />
<br><br><br><br><br><br><br><br><br />
<strong>Nimas Ghasani</strong><br><br />
<strong>Risma Wiharyanti</strong><br><br />
<strong>Bakhtiar Hermawan</strong><br><br />
<strong>Wuddan Nadhirah Rodiana</strong><br>School of Life Sciences and Technology<br><br><br />
<strong>Elfina Marchantia</strong><br>Faculty of Mathematics and Natural Science<br><br><br />
</div><br />
</div> <br />
<div class="contenner"><br />
<h1>Wetlab</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/b/b7/WetLab.JPG" width="270px" height="333px"/></div><br />
<div class="desc"><p>They are “The Wetlabors” (wet lab – labor), some people say it because they always work hard to do the wet lab project. But they never hesitate to ask for help to the other members to do the Super Wet lab Project.</p><br />
<br><br><br><br><br><br><br><br><br><br><br><br><br />
<strong>Tri Ekawati Heryanto</strong><br><br />
<strong>Pande Putu Erawijantari</strong><br>School of Life Sciences and Technology<br><br><br />
</div><br />
</div> <br />
<div class="contenner"><br />
<h1>Wiki</h1><br />
<div class="image"><img src="" width="270px" height="360px" /></div><br />
<div class="desc"><p>Manage, Create and implement wiki's website</p><br />
<br><br><br><br><br><br><br><br><br><br><br><br><br />
<strong>Rizky Kusumah</strong><br><br />
<strong>Muhammad Hariomurti Mardikusumo</strong><br>School of Electrical Engineering and Informatics<br><br><br />
</div><br />
</div> <br />
<div class="contenner"> <br />
<br><br />
<h1>Team Advisor</h1><br />
<div class="image2"><img src="https://static.igem.org/mediawiki/igem.org/6/6a/2013_ITB_Indonesia-adv2.jpg" width="154px" height="205px" /></div><br />
<div class="desc2"><strong>Sony Suhandono, Ph.D</strong><p>Genetics and Molecular Biotechnology Research Group<br><br />
School of Life Sciences and Technology <br><br />
Institut Teknologi Bandung</p></div><br />
</div> <br><br />
<div><br />
<div class="image2"><img src="https://static.igem.org/mediawiki/igem.org/8/83/2013_ITB_Indonesia-adv1.jpg" width="154px" height="205px" /></div><br />
<div class="desc2"><strong>Maelita Ramdani Moeis, Ph.D</strong><p>Genetics and Molecular Biotechnology Research Group<br><br />
School of Life Sciences and Technology <br><br />
Institut Teknologi Bandung</p></div><br />
</div> <br><br />
<div><br />
<div class="image2"><img src="https://static.igem.org/mediawiki/2014/8/8a/Dr_Dessy_Natalia.jpg" width="154px" height="205px" /></div><br />
<div class="desc2"><strong>Dessy Natalia, Ph.D</strong><p>Biochemistry Research Division<br><br />
Faculty of Mathematics and Natural Sciences<br><br />
Institut Teknologi Bandung</p></div><br />
</div> <br><br />
<div><br />
<div class="image2"><img src="https://static.igem.org/mediawiki/2014/9/98/Dr_Mochamad_Apri.jpg" width="154px" height="205px" /></div><br />
<div class="desc2"><strong>Mochamad Apri, Ph.D</strong><p>Industrial and Financial Mathematics Research Division<br><br />
Faculty of Mathematics and Natural Sciences<br><br />
Institut Teknologi Bandung</p></div><br />
</div> <br><br />
<div><br />
<div class="image2"><img src="https://static.igem.org/mediawiki/2014/5/57/Dr_Indra_Wibowo.jpg" width="154px" height="205px" /></div><br />
<div class="desc2"><strong>Indra Wibowo, Ph.D</strong><p>Animal Physiology, Developmentand Biomedical Research Group<br><br />
School of Life Sciences and Technology <br><br />
Institut Teknologi Bandung</p></div><br />
</div> <br />
<h1>Instructor</h1><br />
<div class="image2"><img src="https://static.igem.org/mediawiki/parts/5/52/ITB-aridwijayanti.JPG" width="154px" height="205px" /></div><br />
<div class="desc2"><strong>Ari Dwijayanti, M.Si</strong><p>Genetics and Molecular Biotechnology Research Group<br><br />
School of Life Sciences and Technology <br><br />
Institut Teknologi Bandung</p></div><br />
</div> <br><br />
<br><br />
<br><br />
<br />
<div id="sponsor"></div><br />
</div><br />
</div><br />
</body><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Team
Team:ITB Indonesia/Team
2014-10-17T13:23:37Z
<p>Erawijantari: </p>
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<body><br />
<div id="putih"> <div id="logo-igem" onclick="window.location.href='https://2014.igem.org/'"></div><br />
<div id="center"><br />
<div id="logo"></div><br />
<br />
<div id="header"></div><br />
<br />
<div id="blockNav"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<div class="contenner"><br />
<br />
<h1>Team Leader</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/0/06/Team_Leader_ITB.JPG" width="270px" height="333px" /></div><br />
<div class="desc"><p>The leader of ITB_Indonesia 2014. He keep all of members spirit. Dedicated and enthusiastic to genetic engineering and synthetic biology.</p><br />
<br><br><br><br><br><br><br><br><br><br><br />
<strong>Joko Pebrianto Trinugroho</strong><br>School of Life Science and Technology<br />
</div><br />
</div> <br />
<div class="contenner"><br />
<h1>Public Relation</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/0/0a/Public_Relation_Mardalisa.JPG" width="270px" height="333px" /></div><br />
<div class="desc"><p>Role of public relation in ITB_Indonesia 2014. Public Image strategy, outsearch events, media relations, social media, Handling emergencies.</p><br />
<br><br><br><br><br><br><br><br><br><br />
<strong>Mardalisa</strong><br>School of Life Science and Technology</div><br />
</div> <br />
<div class="contenner"> <br />
<h1>Creative</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/d/d2/Tim_Kreatif.jpg" width="270px" height="360px" /></div><br />
<div class="desc"><p>Most creative peoples in team</p><br />
<br><br><br><br><br><br><br><br><br><br><br />
<strong>Fania Feby Ramadhani</strong><br />
<br><strong>Elfa Norisda Aulianisa</strong><br>School of Life Science and Technology<br />
<br />
<br />
<br><br><strong>Muhammad Musyafa Syahbid</strong><br>School of Architecture, Planning, and Policy Development<br />
<br />
</div><br />
</div> <br />
<div class="contenner"><br />
<h1>Idea Team</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/9/9d/Tim_ide.JPG" width="270px" height="333px" /></div><br />
<div class="desc"><p>Idea Team acts as a mastermind in a construction of a novel device for project of team ITB_Indonesia 2014. Besides that, we also try to create a new improvement for the construction of our novel device and develop a new characterization technique. This improvement is hopefully will increase the efficiency and effectivity of the device and the new characterization technique is expected to minimize the budget of research, easily to carry out, and give more accurate result.</p><br />
<br><br><br><br />
<strong>Tirta Widi Gilang Citradi</strong><br><br />
<strong>Christian Heryakusuma</strong><br>School of Life Sciences and Technology<br><br><br />
<strong>Yovin</strong><br>Faculty of Mathematics and Natural Science<br><br />
</div><br />
</div> <br />
<div class="contenner"><br />
<h1>Sponsorship</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/2/29/Sponsorshipitb.JPG" width="270px" height="338px" /></div><br />
<div class="desc"><p>Acquisition of external fund to cover the project expenses, generate in-kind sponsorships, research corporation that would be interested in out project, prepare and submit excellent sponsorship prorposal. Establish and maintain good relationships with corporate representatives, take responsibility for the day to day fund raising, generate revenue through selling souvenir , food, laboratory lab coat, shirt and pen that can promote to socialized our project.</p><br />
<br><br><br><br />
<strong>Lance Rosa Karo-Karo</strong><br><br />
<strong>Oktira Roka Aji</strong><br><br />
<strong>Kenia Permata Sukma</strong><br><br />
<strong>Wiyudi Gomulya</strong><br><br />
<strong>I Dewa Gde Sathya Deva</strong><br>School of Life Sciences and Technology<br><br><br />
</div><br />
</div> <br />
<div class="contenner"><br />
<h1>Human Practice</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/9/9b/Human_Practice.jpg" width="270px" height="360px"/></div><br />
<div class="desc"><p>We are HUMAN PRACTISE! Theory without us is nothing. We introduce synthetic biology, make collaboration, make project documentation, make synthethic biology become enjoyable science, and spread happiness :)</p><br />
<br><br><br><br><br><br><br><br><br />
<strong>Nimas Ghasani</strong><br><br />
<strong>Risma Wiharyanti</strong><br><br />
<strong>Bakhtiar Hermawan</strong><br><br />
<strong>Wuddan Nadhirah Rodiana</strong><br>School of Life Sciences and Technology<br><br><br />
<strong>Elfina Marchantia</strong><br>Faculty of Mathematics and Natural Science<br><br><br />
</div><br />
</div> <br />
<div class="contenner"><br />
<h1>Wetlab</h1><br />
<div class="image"><img src="https://static.igem.org/mediawiki/2014/b/b7/WetLab.JPG" width="270px" height="333px"/></div><br />
<div class="desc"><p>They are “The Wetlabors” (wet lab – labor), some people say it because they always work hard to do the wet lab project. But they never hesitate to ask for help to the other members to do the Super Wet lab Project.</p><br />
<br><br><br><br><br><br><br><br><br><br><br><br><br />
<strong>Tri Ekawati Heryanto</strong><br><br />
<strong>Pande Putu Erawijantari</strong><br>School of Life Sciences and Technology<br><br><br />
</div><br />
</div> <br />
<div class="contenner"><br />
<h1>Wiki</h1><br />
<div class="image"><img src="" width="270px" height="360px" /></div><br />
<div class="desc"><p>Manage, Create and implement wiki's website</p><br />
<br><br><br><br><br><br><br><br><br><br><br><br><br />
<strong>Rizky Kusumah</strong><br><br />
<strong>Muhammad Hariomurti Mardikusumo</strong><br>School of Electrical Engineering and Informatics<br><br><br />
</div><br />
</div> <br />
<div class="contenner"> <br />
<br><br />
<h1>Team Advisor</h1><br />
<div class="image2"><img src="https://static.igem.org/mediawiki/igem.org/6/6a/2013_ITB_Indonesia-adv2.jpg" width="154px" height="205px" /></div><br />
<div class="desc2"><strong>Sony Suhandono, Ph.D</strong><p>Genetics and Molecular Biotechnology Research Group<br><br />
School of Life Sciences and Technology <br><br />
Institut Teknologi Bandung</p></div><br />
</div> <br><br />
<div><br />
<div class="image2"><img src="https://static.igem.org/mediawiki/igem.org/8/83/2013_ITB_Indonesia-adv1.jpg" width="154px" height="205px" /></div><br />
<div class="desc2"><strong>Maelita Ramdani Moeis, Ph.D</strong><p>Genetics and Molecular Biotechnology Research Group<br><br />
School of Life Sciences and Technology <br><br />
Institut Teknologi Bandung</p></div><br />
</div> <br><br />
<div><br />
<div class="image2"><img src="https://static.igem.org/mediawiki/2014/8/8a/Dr_Dessy_Natalia.jpg" width="154px" height="205px" /></div><br />
<div class="desc2"><strong>Dessy Natalia, Ph.D</strong><p>Biochemistry Research Division<br><br />
Faculty of Mathematics and Natural Sciences<br><br />
Institut Teknologi Bandung</p></div><br />
</div> <br><br />
<div><br />
<div class="image2"><img src="https://static.igem.org/mediawiki/2014/9/98/Dr_Mochamad_Apri.jpg" width="154px" height="205px" /></div><br />
<div class="desc2"><strong>Mochamad Apri, Ph.D</strong><p>Industrial and Financial Mathematics Research Division<br><br />
Faculty of Mathematics and Natural Sciences<br><br />
Institut Teknologi Bandung</p></div><br />
</div> <br><br />
<div><br />
<div class="image2"><img src="https://static.igem.org/mediawiki/2014/5/57/Dr_Indra_Wibowo.jpg" width="154px" height="205px" /></div><br />
<div class="desc2"><strong>Indra Wibowo, Ph.D</strong><p>Animal Physiology, Developmentand Biomedical Research Group<br><br />
School of Life Sciences and Technology <br><br />
Institut Teknologi Bandung</p></div><br />
</div> <br />
<h1>Instructor</h1><br />
<div class="image2"><img src="https://static.igem.org/mediawiki/parts/5/52/ITB-aridwijayanti.JPG" width="154px" height="205px" /></div><br />
<div class="desc2"><strong>Ari Dwijayanti, M.Si</strong><p>Genetics and Molecular Biotechnology Research Group<br><br />
School of Life Sciences and Technology <br><br />
Institut Teknologi Bandung</p></div><br />
</div> <br><br />
<br><br />
<br><br />
<br />
<div id="sponsor"></div><br />
</div><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Parts
Team:ITB Indonesia/Parts
2014-10-17T12:21:31Z
<p>Erawijantari: </p>
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<br />
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<br />
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<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>PARTS</h1><br />
<br><br />
<img src="https://static.igem.org/mediawiki/2014/d/db/Parts1.JPG"><br />
<br><br />
<br><br />
<h1>PARTS DESCRIPTION</h1><br />
<br><br />
<ol type="A"><br />
<li>Reporter Module</li><br />
<img src="https://static.igem.org/mediawiki/2014/8/8c/RepMod.JPG"><br />
<p align="justify">The construction of reporter module is consist of inducible promoter PibpAB <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K339010">(BBa_K339010)</a>, RBS <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, amilCP <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K592025">(BBa_K592025) </a>, and double terminator <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0015"> (Bba_B0015) </a>. BBa_K1387000 consist of RBS (Bba_B0034), amilCP (BBa_K592025) and double terminator (BBa_B0015), while the complete module was contained in <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K1387001">(Bba_K!387001)</a>. The reporter module will have a blue chromoprotein when the inclusion body is formed in the cell. This mechanism then acts as an indicator of cytoplasmic stress, in this situation is caused by inclussion body.</p><br />
<br><li>Degradation Module</li><br />
<img src="https://static.igem.org/mediawiki/2014/6/69/DegMod.JPG"><br />
<p align="justify">The casette of degradation module <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K1387006">(BBa_K1387006) </a> is comprised of constitutive promoter, tet operator, RBS, outer membrane protein A (ompA) fused with LC Cutinase, and double terminator. By using constitutive promoter, we expect the recombinant protein, in this case ompA-LC Cutinase, will be synthesized indefinitely. OmpA is outer membrane protein, it comprises of β–strand B3-B7. Rather than using full sequence of ompA, we use ompA (46-159) fused with lipoprotein signal sequence, thus exposing it on the cell surface of bacteria1. We fuse lpp-ompA with LC Cutinase, an enzyme that exhibit esterase activity. LC-Cutinase esterase activity will cleave the ester bond on PET and releasing terephthalic acid and ethylene glycol as a product. Fusion strategy of lpp-ompA on LC Cutinase gene is one of our way to create a whole cell biocatalyst which will be an effective molecular machinery to degrade PET on the environment. Due to constitutive expression of the recombinant protein in E.coli, there is a possibility of formation of insoluble protein inside the cell (inclusion body), because of that, we put tet operator upstream of RBS for further regulation (Georgiou et al, 1996).</p><br />
<br><li>Self-Regulatory Module</li><br />
<img src="https://static.igem.org/mediawiki/2014/b/b4/Sr1.JPG"><br />
<p align="justify">pIBPAB is a hybrid promoter that induced by inclusion body from unfunctional protein. This promoter followed by strong RBS <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, make a new part called <a style="color:#342D7E" href="http://parts.igem.org/Part:Bba_K1387005"> Bba_K1387005 </a> </p><br />
<img src="https://static.igem.org/mediawiki/2014/d/de/Sr2.JPG"><br />
<p align="justify">This casette <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0034">(BBa_K1387002)</a> consist of TetR <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_C0040">(BBa_C0040)</a> and Double Terminator <a style="color:#342D7E"href="http://parts.igem.org/Part:BBa_B0015"> (Bba_B0015) </a>. TetR is tetracycline repressor that bind to operator region and repressing the expression of downstream gene. In our system, the degradation module consist of TetO (Tet Operator) which is region of TetR (Tet Repressor) placed.</p><br />
<br><img src="https://static.igem.org/mediawiki/2014/f/f2/Sr3.JPG"><br />
<p>The construction of reporter module is consist of inducible promoter PibpAB <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K339010">(BBa_K339010)</a>RBS <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, amilCP <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K592025">(BBa_K592025) </a>, and double terminator <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0015"> (Bba_B0015) </a>. <a style="color:#342D7E"href="hhttp://parts.igem.org/Part:BBa_K1387000"> BBa_K1387000 </a> consist of RBS <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, amilCP <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K592025">(BBa_K592025) </a> and double terminator <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0015"> (Bba_B0015) </a>, while the complete module was contained in <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K1387001">BBa_K1387001 </a>. The reporter module will have a blue chromoprotein when the inclusion body is formed in the cell. This mechanism then acts as an indicator of cytoplasmic stress, in this situation is caused by inclussion body.</p><br />
<br><li>Convertion Module</li><br />
<img src="https://static.igem.org/mediawiki/2014/7/7a/ConMod.JPG"><br />
<p align="justify">Improvisation the part submitted by UC Davis team <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K936024">BBa_K936024</a> by combining the construct with terminator, so we can characterize the expression of the enzymes and performe the assay for ethylene glycol utilization better than before. We name this BioBrick as convertion module.</p><br />
<br><li>References</li><br />
<p>1Georgiou,G., Stephens,L., Stathopoulos,C., Poetschke,L., Mendenhall,J., and Earhart,F. 1996. Display of β-Lactamase on Eschericia coli Surface : Outer Membrane Phenotypes Conferred by Lpp’-ompA’- β-Lactamase Fusions. <i> Protein Engineering, 9, 239-247</i>.</p><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/protocol
Team:ITB Indonesia/protocol
2014-10-17T11:59:45Z
<p>Erawijantari: </p>
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Wetlab Protocol</h1><br />
<ol><br />
<li>SDS PAGE<br><br />
<p align="justify">SDS-PAGE analysis was carried out based on Laemmli System (Laemmli, 1970). First, all of the reagents were mixed together in order to make stacking and separating gel. Bacterial culture was centrifuged at 14.000 g, 1 minutes, then the supernatant was discharged. Pellet cells were mixed with sample buffer then boiled at 100 C for 5 minutes. 10 uL of samples applied to each wells. Electrophoresis perfomed in SDS-Glycine buffer at 130V constant until dye reached the bottom of the gel (Sambrook et al., 2003).</p><br />
</li><br />
<br><br />
<li>Ethylene Glycol Assay using Chromatic Acid<br />
<ol type="a"><br />
<li>Ethylene Glycol Standard Solution Procedure<br />
<ol><br />
<li>Ethylene Glycol concentration ≈ 0.5 M (31035 ppm) -> Dissolve 279 µL ethylene glycol in 10 mL deion water.</li><br />
<li>Ethylene Glycol concentration ≈ 0.05 M (3103.5 ppm) -> Dissolve 300 µL ethylene glycol 0.5 M, then dilute until 3000 µL.</li><br />
<li>Ethylene Glycol concentration ≈ 0.005 M (310.35 ppm) -> Dissolve 300 µL ethylene glycol 0.05 M, then dilute until 3000 µL.</li><br />
<li>Ethylene Glycol concentration ≈ 0.0005 M (31.035 ppm) -> Dissolve 300 µL ethylene glycol 0.005 M, then dilute until 3000 µL.</li><br />
<li>Ethylene Glycol concentration ≈ 0.00005 M (3.1035 ppm) -> Dissolve 300 µL ethylene glycol 0.0005 M, then dilute until 3000 µL.</li><br />
</ol><br />
</li><br />
<li>Chromic Acid Standard Solution Procedure<br />
<ol><br />
<li>Chromic acid ≈ 0.6 M (177000 ppm) -> Dissolve 1.77 gr K2CrO7 (Mr 294.18) with sulphate acid until the volume 10 mL.</li><br />
<li>Do dilution of chromic acid same as ethylene glycol standard solution procedure.</li><br />
</ol><br />
</li><br />
<li>Ethylene Glycol Concentration Measurement through Spectrofotometri UV-Vis<br>4 Cr2O72- + 3 C2H6O2 + 32 H+ -> 8 Cr3+ + 3 C2H2O4 + 22 H2O<br />
<ol><br />
<li>Ethylene glycol 300 ppm -> Pipette 29 µL ethylene glycol solution 31.035 ppm then dilute it until 3 mL.</li><br />
<li>Chromic acid 3.000 ppm -> Pipette 8.5 µL chromic acid solution 177.000 ppm then dilute it until 5 mL.</li><br />
<li>Sulphate acid blank solution -> Enter 1 mL sulphate acid into cuvette, then search maximum wavelength for this solution.</li><br />
<li>Chromic acid blank solution -> Enter 1 mL chromic acid 300 pm into cuvette, then search maximum wavelength for this solution.</li><br />
<li>Add 600 µL chromic acid 300 ppm and 400 µL ethylene glycol, then measure the absorbance of this solution.<br><br />
The difference absorbance values between chromic acid were added with sulfuric acid and ethylene glycol can be used as a parameter to calculate the concentration of ethylene glycol in the sample.</li><br />
</ol><br />
</li><br />
<li>Qualitative and Quantitative Ethylene Glycol Measurement<br>Qualitative Procedure:<br />
<ol><br />
<li>Add 0.25 mL sampel into microtube</li><br />
<li>Add 0.5 mL Acetone and 0.1 mL Chromic acid (H2CrO4) into microtube</li><br />
<li>Mix the mixture</li><br />
<li>Incubate the mixture in waterbath at 65 - 700C for 10 minutes<br><br />
Positive result will show the color changes from orange to green</li><br />
</ol><br />
Quantitative Procedure:<br />
<ol><br />
<li>Add 0.25 mL sampel into microtube</li><br />
<li>Add 0.5 mL Acetone and 0.1 mL Chromic acid (H2CrO4) into microtube</li><br />
<li>Mix the mixture</li><br />
<li>Incubate the mixture in waterbath at 65 - 700C for 10 minutes</li><br />
<li>Check the absorbance at A446 nm</li><br />
</ol><br />
</li><br />
</ol><br />
</li><br />
<br><br />
<li>pNPP Assay<br><br />
<p align="justify">Cutinase activity assays were performed using bacterial culture. The activity was determined using spectrophotometric assay at 410 nm with p-nitrophenyl palmitate (pNPP) as a substrate. pNPP was dissolved in acetonitrile at a concentration of 10 mM. Ethanol and 50 mM Tris-Cl buffer (pH 8.0) were added to a ratio of 1:4:95 (v/v/v) of acetonitrile/ ethanol/ buffer (Lee, 1999). 0.3 mL of the bacterial culture was reacted to the substrate solution (0.9 mL). Then, it was incubated at 25C for 15 minutes then centrifuge at 12,000g, 15 min, and 4C . Enzyme activity was measured by monitoring the change in absorbance at λ 410 nm that represents the amount of released p-nitrophenol (pNP). The activity of cutinase was determined based on the standard curve of p-nitrophenol. One unit of cutinase activity was defined as the amount of enzyme releasing 1 μmol pNP per minute under the assay conditions (Lee et al., 1999).</p><br />
</li><br />
<br><br />
<li>SEM (Scanning Electron Microscope)<br><br />
<p>For biodegrading experiments, we used 3x5 cm2 bottle plastics with average weight 0.05 g. Then, we washed it several times with water and ethanol. Plastic sample was applying just before we inoculated the bacteria on the medium. Bacterial culture were grown at 37C in Luria-Broth medium. After 3 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss.</p><br />
<p>Meanwhile, for UC Davis bacterial culture, we inoculated the bacteria in Luria-Broth medium supplemented with 170 g/ml cloramphenicol. After an absorbance A600 nm of 0.6 was reached, we added 1% of arabinose and plastic sample. After 2 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss. We used medium supplemented with antibiotics and plastic sample as control on this experiment.</p><br />
</li><br />
<br><br />
<li>References<br />
<ol><br />
<li>Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. <i>Nature 227, 680-685.</i></li><br />
<li>Lee, Dong-Woo, Koh,Y.S.,Kim, K.J., Kim, B., et al. 1999. Isolation and characterization of a thermophilic lipase from Bacillus thermoleovorans ID-1, FEMS, Microbiology Letters. <i>179: 393-400.</i></li><br />
<li>Sambrook, J. 2003. <i>Molecular Cloning: a Laboratory Manual</i>. Cold Spring Harbor Laboratory: Cold Spring Harbor.</li><br />
</ol><br />
</li><br />
</ol><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/protocol
Team:ITB Indonesia/protocol
2014-10-17T11:58:29Z
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
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<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
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<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Wetlab Protocol</h1><br />
<ol><br />
<li>SDS PAGE<br><br />
<p align="justify">SDS-PAGE analysis was carried out based on Laemmli System (Laemmli, 1970). First, all of the reagents were mixed together in order to make stacking and separating gel. Bacterial culture was centrifuged at 14.000 g, 1 minutes, then the supernatant was discharged. Pellet cells were mixed with sample buffer then boiled at 100 C for 5 minutes. 10 uL of samples applied to each wells. Electrophoresis perfomed in SDS-Glycine buffer at 130V constant until dye reached the bottom of the gel (Sambrook et al., 2003).</p><br />
</li><br />
<br><br />
<li>Ethylene Glycol Assay using Chromatic Acid<br />
<ol type="a"><br />
<li>Ethylene Glycol Standard Solution Procedure<br />
<ol><br />
<li>Ethylene Glycol concentration ≈ 0.5 M (31035 ppm) -> Dissolve 279 µL ethylene glycol in 10 mL deion water.</li><br />
<li>Ethylene Glycol concentration ≈ 0.05 M (3103.5 ppm) -> Dissolve 300 µL ethylene glycol 0.5 M, then dilute until 3000 µL.</li><br />
<li>Ethylene Glycol concentration ≈ 0.005 M (310.35 ppm) -> Dissolve 300 µL ethylene glycol 0.05 M, then dilute until 3000 µL.</li><br />
<li>Ethylene Glycol concentration ≈ 0.0005 M (31.035 ppm) -> Dissolve 300 µL ethylene glycol 0.005 M, then dilute until 3000 µL.</li><br />
<li>Ethylene Glycol concentration ≈ 0.00005 M (3.1035 ppm) -> Dissolve 300 µL ethylene glycol 0.0005 M, then dilute until 3000 µL.</li><br />
</ol><br />
</li><br />
<li>Chromic Acid Standard Solution Procedure<br />
<ol><br />
<li>Chromic acid ≈ 0.6 M (177000 ppm) -> Dissolve 1.77 gr K2CrO7 (Mr 294.18) with sulphate acid until the volume 10 mL.</li><br />
<li>Do dilution of chromic acid same as ethylene glycol standard solution procedure.</li><br />
</ol><br />
</li><br />
<li>Ethylene Glycol Concentration Measurement through Spectrofotometri UV-Vis<br>4 Cr2O72- + 3 C2H6O2 + 32 H+ -> 8 Cr3+ + 3 C2H2O4 + 22 H2O<br />
<ol><br />
<li>Ethylene glycol 300 ppm -> Pipette 29 µL ethylene glycol solution 31.035 ppm then dilute it until 3 mL.</li><br />
<li>Chromic acid 3.000 ppm -> Pipette 8.5 µL chromic acid solution 177.000 ppm then dilute it until 5 mL.</li><br />
<li>Sulphate acid blank solution -> Enter 1 mL sulphate acid into cuvette, then search maximum wavelength for this solution.</li><br />
<li>Chromic acid blank solution -> Enter 1 mL chromic acid 300 pm into cuvette, then search maximum wavelength for this solution.</li><br />
<li>Add 600 µL chromic acid 300 ppm and 400 µL ethylene glycol, then measure the absorbance of this solution.<br><br />
The difference absorbance values between chromic acid were added with sulfuric acid and ethylene glycol can be used as a parameter to calculate the concentration of ethylene glycol in the sample.</li><br />
</ol><br />
</li><br />
<li>Qualitative and Quantitative Ethylene Glycol Measurement<br>Qualitative Procedure:<br />
<ol><br />
<li>Add 0.25 mL sampel into microtube</li><br />
<li>Add 0.5 mL Acetone and 0.1 mL Chromic acid (H2CrO4) into microtube</li><br />
<li>Mix the mixture</li><br />
<li>Incubate the mixture in waterbath at 65 - 700C for 10 minutes<br><br />
Positive result will show the color changes from orange to green</li><br />
</ol><br />
Quantitative Procedure:<br />
<ol><br />
<li>Add 0.25 mL sampel into microtube</li><br />
<li>Add 0.5 mL Acetone and 0.1 mL Chromic acid (H2CrO4) into microtube</li><br />
<li>Mix the mixture</li><br />
<li>Incubate the mixture in waterbath at 65 - 700C for 10 minutes</li><br />
<li>Check the absorbance at A446 nm</li><br />
</ol><br />
</li><br />
</ol><br />
</li><br />
<br><br />
<li>pNPP Assay<br><br />
<p align="justify">Cutinase activity assays were performed using bacterial culture. The activity was determined using spectrophotometric assay at 410 nm with p-nitrophenyl palmitate (pNPP) as a substrate. pNPP was dissolved in acetonitrile at a concentration of 10 mM. Ethanol and 50 mM Tris-Cl buffer (pH 8.0) were added to a ratio of 1:4:95 (v/v/v) of acetonitrile/ ethanol/ buffer (Lee, 1999). 0.3 mL of the bacterial culture was reacted to the substrate solution (0.9 mL). Then, it was incubated at 25C for 15 minutes then centrifuge at 12,000g, 15 min, and 4C . Enzyme activity was measured by monitoring the change in absorbance at λ 410 nm that represents the amount of released p-nitrophenol (pNP). The activity of cutinase was determined based on the standard curve of p-nitrophenol. One unit of cutinase activity was defined as the amount of enzyme releasing 1 μmol pNP per minute under the assay conditions (Lee et al., 1999).</p><br />
</li><br />
<br><br />
<li>SEM (Scanning Electron Microscope)<br><br />
<p>For biodegrading experiments, we used 3x5 cm2 bottle plastics with average weight 0.05 g. Then, we washed it several times with water and ethanol. Plastic sample was applying just before we inoculated the bacteria on the medium. Bacterial culture were grown at 37C in Luria-Broth medium. After 3 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss.</p><br />
<p>Meanwhile, for UC Davis bacterial culture, we inoculated the bacteria in Luria-Broth medium supplemented with 170 g/ml cloramphenicol. After an absorbance A600 nm of 0.6 was reached, we added 1% of arabinose and plastic sample. After 2 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss. We used medium supplemented with antibiotics and plastic sample as control on this experiment.</p><br />
</li><br />
<br><br />
<li>References<br />
<ol><br />
<li>Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685.</li><br />
<li>Lee, Dong-Woo, Koh,Y.S.,Kim, K.J., Kim, B., et al. 1999. Isolation and characterization of a thermophilic lipase from Bacillus thermoleovorans ID-1, FEMS, Microbiology Letters. 179: 393-400.</li><br />
<li>Sambrook, J. 2003. Molecular Cloning: a Laboratory Manual. Cold Spring Harbor Laboratory: Cold Spring Harbor.</li><br />
</ol><br />
</li><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/protocol
Team:ITB Indonesia/protocol
2014-10-17T11:57:37Z
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<li>TEAM<br />
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<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
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<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
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</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Wetlab Protocol</h1><br />
<ol><br />
<li>SDS PAGE<br><br />
<p align="justify">SDS-PAGE analysis was carried out based on Laemmli System (Laemmli, 1970). First, all of the reagents were mixed together in order to make stacking and separating gel. Bacterial culture was centrifuged at 14.000 g, 1 minutes, then the supernatant was discharged. Pellet cells were mixed with sample buffer then boiled at 100 C for 5 minutes. 10 uL of samples applied to each wells. Electrophoresis perfomed in SDS-Glycine buffer at 130V constant until dye reached the bottom of the gel (Sambrook et al., 2003).</p><br />
</li><br />
<br><br />
<li>Ethylene Glycol Assay using Chromatic Acid<br />
<ol type="a"><br />
<li>Ethylene Glycol Standard Solution Procedure<br />
<ol><br />
<li>Ethylene Glycol concentration ≈ 0.5 M (31035 ppm) -> Dissolve 279 µL ethylene glycol in 10 mL deion water.</li><br />
<li>Ethylene Glycol concentration ≈ 0.05 M (3103.5 ppm) -> Dissolve 300 µL ethylene glycol 0.5 M, then dilute until 3000 µL.</li><br />
<li>Ethylene Glycol concentration ≈ 0.005 M (310.35 ppm) -> Dissolve 300 µL ethylene glycol 0.05 M, then dilute until 3000 µL.</li><br />
<li>Ethylene Glycol concentration ≈ 0.0005 M (31.035 ppm) -> Dissolve 300 µL ethylene glycol 0.005 M, then dilute until 3000 µL.</li><br />
<li>Ethylene Glycol concentration ≈ 0.00005 M (3.1035 ppm) -> Dissolve 300 µL ethylene glycol 0.0005 M, then dilute until 3000 µL.</li><br />
</ol><br />
</li><br />
<li>Chromic Acid Standard Solution Procedure<br />
<ol><br />
<li>Chromic acid ≈ 0.6 M (177000 ppm) -> Dissolve 1.77 gr K2CrO7 (Mr 294.18) with sulphate acid until the volume 10 mL.</li><br />
<li>Do dilution of chromic acid same as ethylene glycol standard solution procedure.</li><br />
</ol><br />
</li><br />
<li>Ethylene Glycol Concentration Measurement through Spectrofotometri UV-Vis<br>4 Cr2O72- + 3 C2H6O2 + 32 H+ -> 8 Cr3+ + 3 C2H2O4 + 22 H2O<br />
<ol><br />
<li>Ethylene glycol 300 ppm -> Pipette 29 µL ethylene glycol solution 31.035 ppm then dilute it until 3 mL.</li><br />
<li>Chromic acid 3.000 ppm -> Pipette 8.5 µL chromic acid solution 177.000 ppm then dilute it until 5 mL.</li><br />
<li>Sulphate acid blank solution -> Enter 1 mL sulphate acid into cuvette, then search maximum wavelength for this solution.</li><br />
<li>Chromic acid blank solution -> Enter 1 mL chromic acid 300 pm into cuvette, then search maximum wavelength for this solution.</li><br />
<li>Add 600 µL chromic acid 300 ppm and 400 µL ethylene glycol, then measure the absorbance of this solution.<br><br />
The difference absorbance values between chromic acid were added with sulfuric acid and ethylene glycol can be used as a parameter to calculate the concentration of ethylene glycol in the sample.</li><br />
</ol><br />
</li><br />
<li>Qualitative and Quantitative Ethylene Glycol Measurement<br>Qualitative Procedure:<br />
<ol><br />
<li>Add 0.25 mL sampel into microtube</li><br />
<li>Add 0.5 mL Acetone and 0.1 mL Chromic acid (H2CrO4) into microtube</li><br />
<li>Mix the mixture</li><br />
<li>Incubate the mixture in waterbath at 65 - 700C for 10 minutes<br><br />
Positive result will show the color changes from orange to green</li><br />
</ol><br />
Quantitative Procedure:<br />
<ol><br />
<li>Add 0.25 mL sampel into microtube</li><br />
<li>Add 0.5 mL Acetone and 0.1 mL Chromic acid (H2CrO4) into microtube</li><br />
<li>Mix the mixture</li><br />
<li>Incubate the mixture in waterbath at 65 - 700C for 10 minutes</li><br />
<li>Check the absorbance at A446 nm</li><br />
</ol><br />
</li><br />
</ol><br />
</li><br />
<br><br />
<li>pNPP Assay<br><br />
<p>Cutinase activity assays were performed using bacterial culture. The activity was determined using spectrophotometric assay at 410 nm with p-nitrophenyl palmitate (pNPP) as a substrate. pNPP was dissolved in acetonitrile at a concentration of 10 mM. Ethanol and 50 mM Tris-Cl buffer (pH 8.0) were added to a ratio of 1:4:95 (v/v/v) of acetonitrile/ ethanol/ buffer (Lee, 1999). 0.3 mL of the bacterial culture was reacted to the substrate solution (0.9 mL). Then, it was incubated at 25C for 15 minutes then centrifuge at 12,000g, 15 min, and 4C . Enzyme activity was measured by monitoring the change in absorbance at λ 410 nm that represents the amount of released p-nitrophenol (pNP). The activity of cutinase was determined based on the standard curve of p-nitrophenol. One unit of cutinase activity was defined as the amount of enzyme releasing 1 μmol pNP per minute under the assay conditions (Lee et al., 1999).</p><br />
</li><br />
<br><br />
<li>SEM (Scanning Electron Microscope)<br><br />
<p>For biodegrading experiments, we used 3x5 cm2 bottle plastics with average weight 0.05 g. Then, we washed it several times with water and ethanol. Plastic sample was applying just before we inoculated the bacteria on the medium. Bacterial culture were grown at 37C in Luria-Broth medium. After 3 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss.</p><br />
<p>Meanwhile, for UC Davis bacterial culture, we inoculated the bacteria in Luria-Broth medium supplemented with 170 g/ml cloramphenicol. After an absorbance A600 nm of 0.6 was reached, we added 1% of arabinose and plastic sample. After 2 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss. We used medium supplemented with antibiotics and plastic sample as control on this experiment.</p><br />
</li><br />
<br><br />
<li>References<br />
<ol><br />
<li>Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685.</li><br />
<li>Lee, Dong-Woo, Koh,Y.S.,Kim, K.J., Kim, B., et al. 1999. Isolation and characterization of a thermophilic lipase from Bacillus thermoleovorans ID-1, FEMS, Microbiology Letters. 179: 393-400.</li><br />
<li>Sambrook, J. 2003. Molecular Cloning: a Laboratory Manual. Cold Spring Harbor Laboratory: Cold Spring Harbor.</li><br />
</ol><br />
</li><br />
</ol><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/protocol
Team:ITB Indonesia/protocol
2014-10-17T08:52:43Z
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
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<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
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<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Wetlab Protocol</h1><br />
<ol><br />
<li>SDS PAGE<br><br />
<p align="justify">SDS-PAGE analysis was carried out based on Laemmli System (Laemmli, 1970). First, all of the reagents were mixed together in order to make stacking and separating gel. Bacterial culture was centrifuged at 14.000 g, 1 minutes, then the supernatant was discharged. Pellet cells were mixed with sample buffer then boiled at 100 C for 5 minutes. 10 uL of samples applied to each wells. Electrophoresis perfomed in SDS-Glycine buffer at 130V constant until dye reached the bottom of the gel (Sambrook et al., 2003).</p><br />
</li><br />
<br><br />
<li>Ethylene Glycol Assay using Chromatic Acid<br />
<ol type="a"><br />
<li>Ethylene Glycol Standard Solution Procedure<br />
<ol><br />
<li>Ethylene Glycol concentration ≈ 0.5 M (31035 ppm) -> Dissolve 279 µL ethylene glycol in 10 mL deion water.</li><br />
<li>Ethylene Glycol concentration ≈ 0.05 M (3103.5 ppm) -> Dissolve 300 µL ethylene glycol 0.5 M, then dilute until 3000 µL.</li><br />
<li>Ethylene Glycol concentration ≈ 0.005 M (310.35 ppm) -> Dissolve 300 µL ethylene glycol 0.05 M, then dilute until 3000 µL.</li><br />
<li>Ethylene Glycol concentration ≈ 0.0005 M (31.035 ppm) -> Dissolve 300 µL ethylene glycol 0.005 M, then dilute until 3000 µL.</li><br />
<li>Ethylene Glycol concentration ≈ 0.00005 M (3.1035 ppm) -> Dissolve 300 µL ethylene glycol 0.0005 M, then dilute until 3000 µL.</li><br />
</ol><br />
</li><br />
<li>Chromic Acid Standard Solution Procedure<br />
<ol><br />
<li>Chromic acid ≈ 0.6 M (177000 ppm) -> Dissolve 1.77 gr K2CrO7 (Mr 294.18) with sulphate acid until the volume 10 mL.</li><br />
<li>Do dilution of chromic acid same as ethylene glycol standard solution procedure.</li><br />
</ol><br />
</li><br />
<li>Ethylene Glycol Concentration Measurement through Spectrofotometri UV-Vis<br>4 Cr2O72- + 3 C2H6O2 + 32 H+ -> 8 Cr3+ + 3 C2H2O4 + 22 H2O<br />
<ol><br />
<li>Ethylene glycol 300 ppm -> Pipette 29 µL ethylene glycol solution 31.035 ppm then dilute it until 3 mL.</li><br />
<li>Chromic acid 3.000 ppm -> Pipette 8.5 µL chromic acid solution 177.000 ppm then dilute it until 5 mL.</li><br />
<li>Sulphate acid blank solution -> Enter 1 mL sulphate acid into cuvette, then search maximum wavelength for this solution.</li><br />
<li>Chromic acid blank solution -> Enter 1 mL chromic acid 300 pm into cuvette, then search maximum wavelength for this solution.</li><br />
<li>Add 600 µL chromic acid 300 ppm and 400 µL ethylene glycol, then measure the absorbance of this solution.<br><br />
The difference absorbance values between chromic acid were added with sulfuric acid and ethylene glycol can be used as a parameter to calculate the concentration of ethylene glycol in the sample.</li><br />
</ol><br />
</li><br />
<li>Qualitative and Quantitative Ethylene Glycol Measurement<br>Qualitative Procedure:<br />
<ol><br />
<li>Add 0.25 mL sampel into microtube</li><br />
<li>Add 0.5 mL Acetone and 0.1 mL Chromic acid (H2CrO4) into microtube</li><br />
<li>Mix the mixture</li><br />
<li>Incubate the mixture in waterbath at 65 - 700C for 10 minutes<br><br />
Positive result will show the color changes from orange to green</li><br />
</ol><br />
Quantitative Procedure:<br />
<ol><br />
<li>Add 0.25 mL sampel into microtube</li><br />
<li>Add 0.5 mL Acetone and 0.1 mL Chromic acid (H2CrO4) into microtube</li><br />
<li>Mix the mixture</li><br />
<li>Incubate the mixture in waterbath at 65 - 700C for 10 minutes</li><br />
<li>Check the absorbance at A446 nm</li><br />
</ol><br />
</li><br />
</ol><br />
</li><br />
<br><br />
<li>pNPP Assay<br><br />
<p>Cutinase activity assays were performed using bacterial culture. The activity was determined using spectrophotometric assay at 410 nm with p-nitrophenyl palmitate (pNPP) as a substrate. pNPP was dissolved in acetonitrile at a concentration of 10 mM. Ethanol and 50 mM Tris-Cl buffer (pH 8.0) were added to a ratio of 1:4:95 (v/v/v) of acetonitrile/ ethanol/ buffer (Lee, 1999). 0.3 mL of the bacterial culture was reacted to the substrate solution (0.9 mL). Then, it was incubated at 25C for 15 minutes then centrifuge at 12,000g, 15 min, and 4C . Enzyme activity was measured by monitoring the change in absorbance at λ 410 nm that represents the amount of released p-nitrophenol (pNP). The activity of cutinase was determined based on the standard curve of p-nitrophenol. One unit of cutinase activity was defined as the amount of enzyme releasing 1 μmol pNP per minute under the assay conditions (Lee et al., 1999).</p><br />
</li><br />
<br><br />
<li>SEM (Scanning Electron Microscope)<br><br />
<p>For biodegrading experiments, we used 3x5 cm2 bottle plastics with average weight 0.05 g. Then, we washed it several times with water and ethanol. Plastic sample was applied just before we inoculated the bacteria on the medium. Bacterial culture were grown at 37C in Luria-Broth medium. After 3 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss.</p><br />
<p>Meanwhile, for UC Davis bacterial culture, we inoculated the bacteria in Luria-Broth medium supplemented with 170 g/ml cloramphenicol. After an absorbance A600 nm of 0.6 was reached, we added 1% of arabinose and plastic sample. After 2 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss. We used medium supplemented with antibiotics and plastic sample as control on this experiment.</p><br />
</li><br />
<br><br />
<li>References<br />
<ol><br />
<li>Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685.</li><br />
<li>Lee, Dong-Woo, Koh,Y.S.,Kim, K.J., Kim, B., et al. 1999. Isolation and characterization of a thermophilic lipase from Bacillus thermoleovorans ID-1, FEMS, Microbiology Letters. 179: 393-400.</li><br />
<li>Sambrook, J. 2003. Molecular Cloning: a Laboratory Manual. Cold Spring Harbor Laboratory: Cold Spring Harbor.</li><br />
</ol><br />
</li><br />
</ol><br />
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http://2014.igem.org/Team:ITB_Indonesia/SelfMod
Team:ITB Indonesia/SelfMod
2014-10-17T03:47:13Z
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<br />
<br><br />
<h1>Self Regulatory Module</h1><br />
<p align="justify">Self-regulatory module is designed to repair the inefficiency of system due to formation of inclusion body. Inclusion body is formed from aggregation of misfolded protein caused by overexpression of LC Cutinase protein (degradation module). Besides being inefficient to the system, inclusion body may also kill the organism because it gives a cytoplasmic stress to cell.</p><br />
<p align="justify">When inclusion body occurs, inclusion body inducible promoter (PibpAB) will active and the expression of TetR repressor will take place. This TetR repressor then corresponds to TetO in degradation module and inhibits the expression of LC Cutinase protein. This inhibition effect will eventually decreasing the inclusion body present in the cell and improving the system efficiency.</p><br />
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Erawijantari
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Team:ITB Indonesia/SelfMod
2014-10-17T03:45:08Z
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Self Regulatory Module</h1><br />
<p>Self-regulatory module is designed to repair the inefficiency of system due to formation of inclusion body. Inclusion body is formed from aggregation of misfolded protein caused by overexpression of LC Cutinase protein (degradation module). Besides being inefficient to the system, inclusion body may also kill the organism because it gives a cytoplasmic stress to cell.</p><br />
<p>When inclusion body occurs, inclusion body inducible promoter (PibpAB) will active and the expression of TetR repressor will take place. This TetR repressor then corresponds to TetO in degradation module and inhibits the expression of LC Cutinase protein. This inhibition effect will eventually decreasing the inclusion body present in the cell and improving the system efficiency.</p><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/a/a6/Self-module.jpg"><br />
<img src="https://static.igem.org/mediawiki/2014/f/fe/Self-module-2.jpg"><br />
<br />
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<div id="center"><br />
<div id="logo"></div><br />
<br />
<div id="header"></div><br />
<br />
<div id="blockNav"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Self Regulatory Module</h1><br />
<p align="justify">Self-regulatory module is designed to repair the inefficiency of system due to formation of inclusion body. Inclusion body is formed from aggregation of misfolded protein caused by overexpression of LC Cutinase protein (degradation module). Besides being inefficient to the system, inclusion body may also kill the organism because it gives a cytoplasmic stress to cell.</p><br />
<p align="justify">When inclusion body occurs, inclusion body inducible promoter (PibpAB) will active and the expression of TetR repressor will take place. This TetR repressor then corresponds to TetO in degradation module and inhibits the expression of LC Cutinase protein. This inhibition effect will eventually decreasing the inclusion body present in the cell and improving the system efficiency.</p><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/a/a6/Self-module.jpg"><br />
<img src="https://static.igem.org/mediawiki/2014/f/fe/Self-module-2.jpg"><br />
<br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/SelfMod
Team:ITB Indonesia/SelfMod
2014-10-17T03:43:52Z
<p>Erawijantari: </p>
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<body><br />
<div id="putih"><div id="logo-igem" onclick="window.location.href='https://2014.igem.org/'"></div><br />
<div id="center"><br />
<div id="logo"></div><br />
<br />
<div id="header"></div><br />
<br />
<div id="blockNav"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Self Regulatory Module</h1><br />
<p align="justify">Self-regulatory module is designed to repair the inefficiency of system due to formation of inclusion body. Inclusion body is formed from aggregation of misfolded protein caused by overexpression of LC Cutinase protein (degradation module). Besides being inefficient to the system, inclusion body may also kill the organism because it gives a cytoplasmic stress to cell.</p><br />
<p align="justify">When inclusion body occurs, inclusion body inducible promoter (PibpAB) will active and the expression of TetR repressor will take place. This TetR repressor then corresponds to TetO in degradation module and inhibits the expression of LC Cutinase protein. This inhibition effect will eventually decreasing the inclusion body present in the cell and improving the system efficiency.</p><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/a/a6/Self-module.jpg"><br />
<img src="https://static.igem.org/mediawiki/2014/f/fe/Self-module-2.jpg"><br />
<br />
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<div id="sponsor"></div><br />
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<div id="center"><br />
<div id="logo"></div><br />
<br />
<div id="header"></div><br />
<br />
<div id="blockNav"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Self Regulatory Module</h1><br />
<p align="justify">Self-regulatory module is designed to repair the inefficiency of system due to formation of inclusion body. Inclusion body is formed from aggregation of misfolded protein caused by overexpression of LC Cutinase protein (degradation module). Besides being inefficient to the system, inclusion body may also kill the organism because it gives a cytoplasmic stress to cell.</p><br />
<p align="justify">When inclusion body occurs, inclusion body inducible promoter (PibpAB) will active and the expression of TetR repressor will take place. This TetR repressor then corresponds to TetO in degradation module and inhibits the expression of LC Cutinase protein. This inhibition effect will eventually decreasing the inclusion body present in the cell and improving the system efficiency.</p><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/a/a6/Self-module.jpg"><br />
<img src="https://static.igem.org/mediawiki/2014/f/fe/Self-module-2.jpg"><br />
<br />
<br><br />
<br><br />
<br />
<div id="sponsor"></div><br />
</div><br />
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</body><br />
</html></div>
Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/SelfMod
Team:ITB Indonesia/SelfMod
2014-10-17T03:42:11Z
<p>Erawijantari: </p>
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<div id="putih"><div id="logo-igem" onclick="window.location.href='https://2014.igem.org/'"></div><br />
<div id="center"><br />
<div id="logo"></div><br />
<br />
<div id="header"></div><br />
<br />
<div id="blockNav"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Self Regulatory Module</h1><br />
<p>Self-regulatory module is designed to repair the inefficiency of system due to formation of inclusion body. Inclusion body is formed from aggregation of misfolded protein caused by overexpression of LC Cutinase protein (degradation module). Besides being inefficient to the system, inclusion body may also kill the organism because it gives a cytoplasmic stress to cell.</p><br />
<p>When inclusion body occurs, inclusion body inducible promoter (PibpAB) will active and the expression of TetR repressor will take place. This TetR repressor then corresponds to TetO in degradation module and inhibits the expression of LC Cutinase protein. This inhibition effect will eventually decreasing the inclusion body present in the cell and improving the system efficiency.</p><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/a/a6/Self-module.jpg"><br />
<img src="https://static.igem.org/mediawiki/2014/f/fe/Self-module-2.jpg"><br />
<br />
<br><br />
<br><br />
<br />
<div id="sponsor"></div><br />
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<div id="center"><br />
<div id="logo"></div><br />
<br />
<div id="header"></div><br />
<br />
<div id="blockNav"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Self Regulatory Module</h1><br />
<p align="justify">Self-regulatory module is designed to repair the inefficiency of system due to formation of inclusion body. Inclusion body is formed from aggregation of misfolded protein caused by overexpression of LC Cutinase protein (degradation module). Besides being inefficient to the system, inclusion body may also kill the organism because it gives a cytoplasmic stress to cell.</p><br />
<p align="justify">When inclusion body occurs, inclusion body inducible promoter (PibpAB) will active and the expression of TetR repressor will take place. This TetR repressor then corresponds to TetO in degradation module and inhibits the expression of LC Cutinase protein. This inhibition effect will eventually decreasing the inclusion body present in the cell and improving the system efficiency.</p><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/a/a6/Self-module.jpg"><br />
<img src="https://static.igem.org/mediawiki/2014/f/fe/Self-module-2.jpg"><br />
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<br><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/SelfMod
Team:ITB Indonesia/SelfMod
2014-10-17T03:41:06Z
<p>Erawijantari: </p>
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<div id="center"><br />
<div id="logo"></div><br />
<br />
<div id="header"></div><br />
<br />
<div id="blockNav"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Self Regulatory Module</h1><br />
<p>Self-regulatory module is designed to repair the inefficiency of system due to formation of inclusion body. Inclusion body is formed from aggregation of misfolded protein caused by overexpression of LC Cutinase protein (degradation module). Besides being inefficient to the system, inclusion body may also kill the organism because it gives a cytoplasmic stress to cell.</p><br />
<p>When inclusion body occurs, inclusion body inducible promoter (PibpAB) will active and the expression of TetR repressor will take place. This TetR repressor then corresponds to TetO in degradation module and inhibits the expression of LC Cutinase protein. This inhibition effect will eventually decreasing the inclusion body present in the cell and improving the system efficiency.</p><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/a/a6/Self-module.jpg"><br />
<img src="https://static.igem.org/mediawiki/2014/f/fe/Self-module-2.jpg"><br />
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<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
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<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
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</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Self Regulatory Module</h1><br />
<p align="justify">Self-regulatory module is designed to repair the inefficiency of system due to formation of inclusion body. Inclusion body is formed from aggregation of misfolded protein caused by overexpression of LC Cutinase protein (degradation module). Besides being inefficient to the system, inclusion body may also kill the organism because it gives a cytoplasmic stress to cell.</p><br />
<p>When inclusion body occurs, inclusion body inducible promoter (PibpAB) will active and the expression of TetR repressor will take place. This TetR repressor then corresponds to TetO in degradation module and inhibits the expression of LC Cutinase protein. This inhibition effect will eventually decreasing the inclusion body present in the cell and improving the system efficiency.</p><br />
<br />
<img src="https://static.igem.org/mediawiki/2014/a/a6/Self-module.jpg"><br />
<img src="https://static.igem.org/mediawiki/2014/f/fe/Self-module-2.jpg"><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/RepMod
Team:ITB Indonesia/RepMod
2014-10-17T03:40:43Z
<p>Erawijantari: </p>
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<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Reporter Module</h1><br />
<p>Reporter module is designed to detect the existence of cytoplasmic stress by naked eye. One of the cytoplasmic stresses is the formation of inclusion body due to overexpression of LC Cutinase in degradation module. The overexpression of the protein may result in an inefficient protein folding where not 100% proteins are useable and probably, the misfolded protein will create an aggregate known as inclusion body that decreasing the system efficiency.</p><br />
<p>Inclusion body inducible promoter (PibpAB) then will detect the formation of inclusion body and report it by expressing amilCP, a blue fluorescent protein that detectable by naked eyes, make it easier to analyze whether the stress level of the cell is in the limit or beyond the limit.</p><br />
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<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Reporter Module</h1><br />
<p align="justify">Reporter module is designed to detect the existence of cytoplasmic stress by naked eye. One of the cytoplasmic stresses is the formation of inclusion body due to overexpression of LC Cutinase in degradation module. The overexpression of the protein may result in an inefficient protein folding where not 100% proteins are useable and probably, the misfolded protein will create an aggregate known as inclusion body that decreasing the system efficiency.</p><br />
<p align="justify">Inclusion body inducible promoter (PibpAB) then will detect the formation of inclusion body and report it by expressing amilCP, a blue fluorescent protein that detectable by naked eyes, make it easier to analyze whether the stress level of the cell is in the limit or beyond the limit.</p><br />
<br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Project
Team:ITB Indonesia/Project
2014-10-17T03:40:09Z
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<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Project Overview</h1><br />
<p align="justify">Team ITB_Indonesia tries to create a new method to degrade plastic using synthetic biology approach. We create a novel device which is a synthetic bacteria that able to degrade PET and consume ethylene glycol as the byproduct of degradation reaction. We also establish ompA protein fused with LC Cutinase in <i>E.coli</i> BL21(DE3) thus exposing the enzymes on the surface membrane of bacteria. This condition make the degradation process easily because PET, a high molecular substrate, doesn't need to pass through bacteria cell membrane. We also construct a cassette comprised of two enzyme, glycoaldehyde reductase and glycoaldehyde dehydrogenase which responsible in metabolising ethylene glycol. We use constitutive promoter for both construct, thus making it necessary to create a modul of reporter and self regulatory in order to maintain system efficiency and minimizing severe cytoplasmic stress, in this case, formation of inclusion body aggregate due to strong constitutive expression.</p><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Parts
Team:ITB Indonesia/Parts
2014-10-17T03:38:52Z
<p>Erawijantari: </p>
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<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>PARTS</h1><br />
<br><br />
<img src="https://static.igem.org/mediawiki/2014/d/db/Parts1.JPG"><br />
<br><br />
<br><br />
<h1>PARTS DESCRIPTION</h1><br />
<br><br />
<ol type="A"><br />
<li>Reporter Module</li><br />
<img src="https://static.igem.org/mediawiki/2014/8/8c/RepMod.JPG"><br />
<p>The construction of reporter module is consist of inducible promoter PibpAB <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K339010">(BBa_K339010)</a>, RBS <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, amilCP <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K592025">(BBa_K592025) </a>, and double terminator <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0015"> (Bba_B0015) </a>. BBa_K1387000 consist of RBS (Bba_B0034), amilCP (BBa_K592025) and double terminator (BBa_B0015), while the complete module was contained in <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K1387001">(Bba_K!387001)</a>. The reporter module will have a blue chromoprotein when the inclusion body is formed in the cell. This mechanism then acts as an indicator of cytoplasmic stress, in this situation is caused by inclussion body.</p><br />
<br><li>Degradation Module</li><br />
<img src="https://static.igem.org/mediawiki/2014/6/69/DegMod.JPG"><br />
<p>The casette of degradation module <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K1387006">(BBa_K1387006) </a> is comprised of constitutive promoter, tet operator, RBS, outer membrane protein A (ompA) fused with LC Cutinase, and double terminator. By using constitutive promoter, we expect the recombinant protein, in this case ompA-LC Cutinase, will be synthesized indefinitely. OmpA is outer membrane protein, it comprises of β–strand B3-B7. Rather than using full sequence of ompA, we use ompA (46-159) fused with lipoprotein signal sequence, thus exposing it on the cell surface of bacteria1. We fuse lpp-ompA with LC Cutinase, an enzyme that exhibit esterase activity. LC-Cutinase esterase activity will cleave the ester bond on PET and releasing terephthalic acid and ethylene glycol as a product. Fusion strategy of lpp-ompA on LC Cutinase gene is one of our way to create a whole cell biocatalyst which will be an effective molecular machinery to degrade PET on the environment. Due to constitutive expression of the recombinant protein in E.coli, there is a possibility of formation of insoluble protein inside the cell (inclusion body), because of that, we put tet operator upstream of RBS for further regulation (Georgiou et al, 1996).</p><br />
<br><li>Self-Regulatory Module</li><br />
<img src="https://static.igem.org/mediawiki/2014/b/b4/Sr1.JPG"><br />
<p>pIBPAB is a hybrid promoter that induced by inclusion body from unfunctional protein. This promoter followed by strong RBS <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, make a new part called </p><br />
<img src="https://static.igem.org/mediawiki/2014/d/de/Sr2.JPG"><br />
<p>This casette <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0034">(BBa_K1387002)</a> consist of TetR <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_C0040">(BBa_C0040)</a> and Double Terminator <a style="color:#342D7E"href="http://parts.igem.org/Part:BBa_B0015"> (Bba_B0015) </a>. TetR is tetracycline repressor that bind to operator region and repressing the expression of downstream gene. In our system, the degradation module consist of TetO (Tet Operator) which is region of TetR (Tet Repressor) placed.</p><br />
<br><img src="https://static.igem.org/mediawiki/2014/f/f2/Sr3.JPG"><br />
<p>The construction of reporter module is consist of inducible promoter PibpAB <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K339010">(BBa_K339010)</a>RBS <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, amilCP <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K592025">(BBa_K592025) </a>, and double terminator <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0015"> (Bba_B0015) </a>. <a style="color:#342D7E"href="hhttp://parts.igem.org/Part:BBa_K1387000"> BBa_K1387000 </a> consist of RBS <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, amilCP <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K592025">(BBa_K592025) </a> and double terminator <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_B0015"> (Bba_B0015) </a>, while the complete module was contained in <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K1387001">BBa_K1387001 </a>. The reporter module will have a blue chromoprotein when the inclusion body is formed in the cell. This mechanism then acts as an indicator of cytoplasmic stress, in this situation is caused by inclussion body.</p><br />
<br><li>Convertion Module</li><br />
<img src="https://static.igem.org/mediawiki/2014/7/7a/ConMod.JPG"><br />
<p>Improvisation the part submitted by UC Davis team <a style="color:#342D7E" href="http://parts.igem.org/Part:BBa_K936024">BBa_K936024</a> by combining the construct with terminator, so we can characterize the expression of the enzymes and performe the assay for ethylene glycol utilization better than before. We name this BioBrick as convertion module.</p><br />
<br><li>References</li><br />
<p>1Georgiou,G., Stephens,L., Stathopoulos,C., Poetschke,L., Mendenhall,J., and Earhart,F. 1996. Display of β-Lactamase on Eschericia coli Surface : Outer Membrane Phenotypes Conferred by Lpp’-ompA’- β-Lactamase Fusions. <i> Protein Engineering, 9, 239-247</i>.</p><br />
</ol><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Parts
Team:ITB Indonesia/Parts
2014-10-17T03:22:22Z
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<br />
<div id="blockNav"><br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>PARTS</h1><br />
<br><br />
<img src="https://static.igem.org/mediawiki/2014/d/db/Parts1.JPG"><br />
<br><br />
<br><br />
<h1>PARTS DESCRIPTION</h1><br />
<br><br />
<ol type="A"><br />
<li>Reporter Module</li><br />
<img src="https://static.igem.org/mediawiki/2014/8/8c/RepMod.JPG"><br />
<p>The construction of reporter module is consist of inducible promoter PibpAB <a style="blue" href="http://parts.igem.org/Part:BBa_K339010">(BBa_K339010)</a>, RBS <a style="blue" href="http://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, amilCP <a style="blue" href="http://parts.igem.org/Part:BBa_K592025">(BBa_K592025) </a>, and double terminator <a style="blue" href="http://parts.igem.org/Part:BBa_B0015"> (Bba_B0015) </a>. BBa_K1387000 consist of RBS (Bba_B0034), amilCP (BBa_K592025) and double terminator (BBa_B0015), while the complete module was contained in <a style="blue" href="http://parts.igem.org/Part:BBa_K1387001">(Bba_K!387001)</a>. The reporter module will have a blue chromoprotein when the inclusion body is formed in the cell. This mechanism then acts as an indicator of cytoplasmic stress, in this situation is caused by inclussion body.</p><br />
<br><li>Degradation Module</li><br />
<img src="https://static.igem.org/mediawiki/2014/6/69/DegMod.JPG"><br />
<p>The casette of degradation module <a style="blue" href="http://parts.igem.org/Part:BBa_K1387006">(BBa_K1387006) </a> is comprised of constitutive promoter, tet operator, RBS, outer membrane protein A (ompA) fused with LC Cutinase, and double terminator. By using constitutive promoter, we expect the recombinant protein, in this case ompA-LC Cutinase, will be synthesized indefinitely. OmpA is outer membrane protein, it comprises of β–strand B3-B7. Rather than using full sequence of ompA, we use ompA (46-159) fused with lipoprotein signal sequence, thus exposing it on the cell surface of bacteria1. We fuse lpp-ompA with LC Cutinase, an enzyme that exhibit esterase activity. LC-Cutinase esterase activity will cleave the ester bond on PET and releasing terephthalic acid and ethylene glycol as a product. Fusion strategy of lpp-ompA on LC Cutinase gene is one of our way to create a whole cell biocatalyst which will be an effective molecular machinery to degrade PET on the environment. Due to constitutive expression of the recombinant protein in E.coli, there is a possibility of formation of insoluble protein inside the cell (inclusion body), because of that, we put tet operator upstream of RBS for further regulation (Georgiou et al, 1996).</p><br />
<br><li>Self-Regulatory Module</li><br />
<img src="https://static.igem.org/mediawiki/2014/b/b4/Sr1.JPG"><br />
<p>pIBPAB is a hybrid promoter that induced by inclusion body from unfunctional protein. This promoter followed by strong RBS <a style="blue" href="http://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, make a new part called </p><br />
<img src="https://static.igem.org/mediawiki/2014/d/de/Sr2.JPG"><br />
<p>This casette <a style="blue" href="http://parts.igem.org/Part:BBa_B0034">(BBa_K1387002)</a> consist of TetR <a style="blue" href="http://parts.igem.org/Part:BBa_C0040">(BBa_C0040)</a> and Double Terminator <a style="blue" href="http://parts.igem.org/Part:BBa_B0015"> (Bba_B0015) </a>. TetR is tetracycline repressor that bind to operator region and repressing the expression of downstream gene. In our system, the degradation module consist of TetO (Tet Operator) which is region of TetR (Tet Repressor) placed.</p><br />
<br><img src="https://static.igem.org/mediawiki/2014/f/f2/Sr3.JPG"><br />
<p>The construction of reporter module is consist of inducible promoter PibpAB <a style="blue" href="http://parts.igem.org/Part:BBa_K339010">(BBa_K339010)</a>(, RBS <a style="blue" href="http://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, amilCP <a style="blue" href="http://parts.igem.org/Part:BBa_K592025">(BBa_K592025) </a>, and double terminator <a style="blue" href="http://parts.igem.org/Part:BBa_B0015"> (Bba_B0015) </a>. <a style="blue" href="hhttp://parts.igem.org/Part:BBa_K1387000"> BBa_K1387000 </a> consist of RBS <a style="blue" href="http://parts.igem.org/Part:BBa_B0034">(BBa_B0034)</a>, amilCP <a style="blue" href="http://parts.igem.org/Part:BBa_K592025">(BBa_K592025) </a> and double terminator <a style="blue" href="http://parts.igem.org/Part:BBa_B0015"> (Bba_B0015) </a>, while the complete module was contained in <a style="blue" href="http://parts.igem.org/Part:BBa_K1387001">BBa_K1387001 </a>. The reporter module will have a blue chromoprotein when the inclusion body is formed in the cell. This mechanism then acts as an indicator of cytoplasmic stress, in this situation is caused by inclussion body.</p><br />
<br><li>Convertion Module</li><br />
<img src="https://static.igem.org/mediawiki/2014/7/7a/ConMod.JPG"><br />
<p>Improvisation the part submitted by UC Davis team <a style="blue" href="http://parts.igem.org/Part:BBa_K936024">BBa_K936024</a> by combining the construct with terminator, so we can characterize the expression of the enzymes and performe the assay for ethylene glycol utilization better than before. We name this BioBrick as convertion module.</p><br />
<br><li>References</li><br />
<p>1Georgiou,G., Stephens,L., Stathopoulos,C., Poetschke,L., Mendenhall,J., and Earhart,F. 1996. Display of β-Lactamase on Eschericia coli Surface : Outer Membrane Phenotypes Conferred by Lpp’-ompA’- β-Lactamase Fusions. Protein Engineering, 9, 239-247.</p><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Modeling
Team:ITB Indonesia/Modeling
2014-10-17T03:19:48Z
<p>Erawijantari: </p>
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
<ul><br />
<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
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<li>PROJECT<br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
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<li>NOTEBOOK<br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
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</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Degradation Module</h1><br />
<p>The degradation reaction of PET<br>PET+LC Cutinase→Ethylene Glycole +Terephtalic Acid+Cutinase</p><br />
<br />
<ol><br />
<li>Cutinase<br><br />
<p align="justify">The amount of LC-Cutinase enzyme is due to the enzyme production rate by <i>Escherichia coli</i> and the degradation of LC-Cutinase. The enzyme (LC-Cutinase) production occurs constitutively (did not need any activator and produced continuously). At the t-time when LC-Cutinase reach the maximum quantity, the Escherichia coli productivity will be decreased because of Inclusion body. Inclusion body is protein which is produced by miss-folded protein. The amount of inclusion body will be increases. When the inclusion body reach the K quantity, it will be repress the LC-Cutinase production because the inclusion body become a inducer for PibPab promoter that could drive Tet Represor gene expression. Our degradation module contain Tet Operator that could be repressed by Tet Represor. Our PibPab promoter also could drive the Amil Cp gene expression that release blue chromoprotein. Based on that phenomenon, the production of LC-Cutinase was modeled using Hill Function with the repressor factor.</p><br />
<img src="https://static.igem.org/mediawiki/parts/e/e4/ITB-DegModellingJPG.JPG"><br />
<br><br />
</li><br />
<li>Inclusion Body<br><br />
<p align="justify">When the <i>Escherichia coli</i> start to produce many of cutinase enzyme at maximum concentration, it start become stress and begin to producing a miss-folded protein called inclusion body. As long as the producing of enzyme cutinase in this condition, the producing of inclusion body will be much. At this time the inclusion body act as the repressor that stop the production of cutinase and also will be the activator of reporter gen (Amil CP). This activity shown by the picture above. The model expression of inclusion body was modeled using Hill Function.</p><br />
<p>The assumption of the model:<br><br />
We do not include the factor of cell division of bacteria<br>Cutinase</p><br />
<img src="https://static.igem.org/mediawiki/2014/e/e4/Mod2.JPG"><br><br />
<p>Inclusion Body inducer</p><br />
<img src="https://static.igem.org/mediawiki/2014/2/2d/Mod3.JPG"><br><br />
<p>Reporter gen (Amil CP) expression</p><br />
<img src="https://static.igem.org/mediawiki/2014/b/b9/Mod4.JPG"><br><br />
<p>Ethylene Glycole amount (as a product of PET degradation)</p><br />
<img src="https://static.igem.org/mediawiki/2014/2/2f/Mod5.JPG"><br><br />
<p>PET degradation</p><br />
<img src="https://static.igem.org/mediawiki/parts/0/08/ITB-Parameters.JPG"><br><br><br />
<p>Parameters</p><br />
<img src="https://static.igem.org/mediawiki/2014/a/a5/Mod6.JPG"><br><br><br />
<p align="justify">Figure 1 shown the graph of cutinase production, inclussion body production and blue protein expression. Figure 2 shown the the close up graph and Figure 3 shown the PET degradation and ethylene glicol product profil by the time of gene expression. Inclussion bodies are reflactile aggreates of protease-resistant misfolded protein that often occur in recombinant bacteria upon gratutious overexpression of cloned genes. In biotechnology, the formation of Ibs represents a main obstacle for protein production since even favouring high protein yields. Aggregation of recombinant proteins is probably due to limiting amount of chaperones when recombinant gene expression is directed at high leves. When the ammount of protein production is maximal, we assume that inclusion body will be release. Inclussion body could drive the expression of Tet represor and AmilCp as shown in figure 1. When the inclussion body could be degraded by the cell, the PibPab promoter will be off and there are no TetR and amilCP expression. In that condition we assume that the the cutinase expression will be increassing until reach some value that could release the inclusion body, than the expression will be represed. In figure 2, we could observe the osilation graph. From the figure 3, we could see that by the time of PET degradation the ethylene glicol product that could be released is increase.</p><br><br />
<img src="https://static.igem.org/mediawiki/2014/6/66/Mod9.JPG"><br><br />
<img src="https://static.igem.org/mediawiki/2014/5/5c/Mod10.JPG"><br><br />
</li><br />
<p<br />
<br><br />
<br />
<li>References<br />
<ol><br />
<li>Carrio, M.M and A. Villaverde. 2002. Construction and Deconstruction of Bacterial Inclusion Bodies. <i>Journal of Biotechnology 96. 3-12</i></li><br />
<li>Chapman, and Halu/CRC, “AN INTRODUCTION TO SYSTEMS BIOLOGY Design Principle of Biological Circuit”</li><br />
<li>Fink, Anthony L. 1998. Protein Aggregation: folding aggregates, inclusion bodies and amyloid. <i>© Current Biology Ltd ISSN 1359-0278</i></li><br />
</ol><br />
</li><br />
</ol><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia
Team:ITB Indonesia
2014-10-17T03:17:34Z
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>What is PET?</h1><br />
<p>PET or polyethylene terephthalate is a thermoplastic polymer resin of the polyester family. This plastic has a characteristic of undegradable by bacteria. Overproduction of this plastic may result in polluting the environment and cause a great impact for any living things that live in the environment.</p><br />
<br><br />
<br />
<h1>Why we must degrade plastic?</h1><br />
<p>As we know, plastic takes 500 or even 1000 years to degrade by process called photodegradation rather than biodegradation. Knowing this fact, it is necessary to degrade the plastic to reduce the quantity of plastic in the environment. Therefore, a brilliant, effective, and efficient degradation method is important, otherwise, the plastic itself will threatening the environment and affected the life of animal and human that lives in there.</p><br />
<br><br />
<br />
<h1>How ColiPlasTer work?</h1><br />
<p>ColiPlasTer is <i>Escherichia coli</i> plastic terminator. These bacteria have a capability to degrade plastic by producing an esterase enzyme that fastens plastic degradation process. This enzyme will degrade plastic to its monomer and using it as a carbon source to sustain the bacteria's life. Therefore, no pollutants produce as a result of this degradation process. Click <a href="https://static.igem.org/mediawiki/2014/c/c8/ITB_INDONESIA_16MB.mov"><font color="#38AF89"><strong>HERE</strong></a> <font color="black">for more detail.</p><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/FutureSystem
Team:ITB Indonesia/FutureSystem
2014-10-17T03:17:07Z
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>CHAPERONE PROTEIN</h1><br />
<p>A misfolded protein may form from aggregation of primary structure of protein that is overexpressed in the cell. This aggregation may create inclusion body which will lower the bacteria metabolic efficiency. If this condition is continuously happened, the death of the cell may occur. Therefore, it is important to decrease the inclusion body in the cell to increase the lifespan of the cell.</p><br />
<p>If chaperone is introduced to the system, it will act as an agent to refold the misfolded protein. This chaperone will be integrated in bacteria genome. The inclusion body then will be refolded and restore the activity of the protein as it normally would. With this system, the efficiency and effectiveness of bacteria recombinant gene expression and metabolic load will be better.</p><br />
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<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
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<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>CHAPERONE PROTEIN</h1><br />
<p align="justify">A misfolded protein may form from aggregation of primary structure of protein that is overexpressed in the cell. This aggregation may create inclusion body which will lower the bacteria metabolic efficiency. If this condition is continuously happened, the death of the cell may occur. Therefore, it is important to decrease the inclusion body in the cell to increase the lifespan of the cell.</p><br />
<p align="justify">If chaperone is introduced to the system, it will act as an agent to refold the misfolded protein. This chaperone will be integrated in bacteria genome. The inclusion body then will be refolded and restore the activity of the protein as it normally would. With this system, the efficiency and effectiveness of bacteria recombinant gene expression and metabolic load will be better.</p><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/FutureSystem
Team:ITB Indonesia/FutureSystem
2014-10-17T03:16:35Z
<p>Erawijantari: </p>
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
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<br />
<br><br />
<h1>CHAPERONE PROTEIN</h1><br />
<p>Amisfolded protein may form from aggregation of primary structure of protein that is overexpressed in the cell. This aggregation may create inclusion body which will lower the bacteria metabolic efficiency. If this condition is continuously happened, the death of the cell may occur. Therefore, it is important to decrease the inclusion body in the cell to increase the lifespan of the cell.</p><br />
<p>If chaperone is introduced to the system, it will act as an agent to refold the misfolded protein. This chaperone will be integrated in bacteria genome. The inclusion body then will be refolded and restore the activity of the protein as it normally would. With this system, the efficiency and effectiveness of bacteria recombinant gene expression and metabolic load will be better.</p><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
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</ul><br />
</div><br />
<br />
<br><br />
<h1>CHAPERONE PROTEIN</h1><br />
<p align="justify">A misfolded protein may form from aggregation of primary structure of protein that is overexpressed in the cell. This aggregation may create inclusion body which will lower the bacteria metabolic efficiency. If this condition is continuously happened, the death of the cell may occur. Therefore, it is important to decrease the inclusion body in the cell to increase the lifespan of the cell.</p><br />
<p align="justify">If chaperone is introduced to the system, it will act as an agent to refold the misfolded protein. This chaperone will be integrated in bacteria genome. The inclusion body then will be refolded and restore the activity of the protein as it normally would. With this system, the efficiency and effectiveness of bacteria recombinant gene expression and metabolic load will be better.</p><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/DegMod
Team:ITB Indonesia/DegMod
2014-10-17T03:15:50Z
<p>Erawijantari: </p>
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
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<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
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<ul><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
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<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Degradation Module</h1><br />
<p align="justify">Team ITB_Indonesia tries to make a module to degrade PET. We use LC-Cutinase fused with ompA and Lpp signal sequence. LC-Cutinase will break the ester bonds on PET structure through esterase activity of the enzyme and generate ethylene glycol and terephthalic acid as product. We plan to expose this enzyme on the surface membrane of bacteria thus making the enzyme immobile and more thermostable. On the other hand, PET which has high molecular mass can be degraded directly without need to pass through bacteria cell membrane. This molecular machine then named as whole cell biocatalyst.</p><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/ConMod
Team:ITB Indonesia/ConMod
2014-10-17T03:14:48Z
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<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Convertion Module</h1><br />
<p align="justify">Convertion module is constructed to create an <i>E.coli</i> that able to survive the present of toxic agent, such as ethylene glycol that being introduce in the system. The module consist of two enzymes, glycoaldehyde reductase and glycoaldehyde dehydrogenase. The enzymes are responsible in metabolising ethylene glycol in <i>E.coli</i> strain BL21 (DE3). The first mentioned enzyme is responsible to catalyse convertion reaction of ethylene glycol to glycoaldehyde while the second one is responsible to catalyse the reaction of glycoaldehyde to glycolate. This intermediate will proceed the next step of metabolic pathway of E.coli and finally ends up on TCA cycle. This process will create an energy (ATP) that can be used by bacteria as carbon source, thus assuring us that ethylene glycol will not pollute the environment.</p><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Attributions
Team:ITB Indonesia/Attributions
2014-10-17T03:13:47Z
<p>Erawijantari: </p>
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
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<li>NOTEBOOK<br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
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<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
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</div><br />
<br />
<br><br />
<p>This project is done iGEM ITB_Indonesia 2014 team members and our collaborator . Our team have worked well dan most dedicated for this project.</p><br />
<h1>Project name: SynBIGreen (Synthetic Biology Indonesia Go Green)</h1><br />
<p>Our team consist of variant background of knowledge, they are from science and engineering. We have many talents that no have one for the others. For finishing this project we mapped our talent to variant job following as:</p><br />
<br />
<h3>Team Leader :</h3><br />
<p>Joko Pebrianto Trinugroho</p><br />
<br />
<h3>Construct :</h3><br />
<p>Tri Ekawati Heryanto, Pande Putu Erawijantari, Oktira Roka Aji, Joko Pebrianto Trinugroho, Nimas Ghassani</p><br />
<br />
<h3>Modeling System :</h3><br />
<p>Aditya Putra Pratama</p><br />
<br />
<h3>Sponsorship team :</h3><br />
<p>JLance Rosa Karo-karo, Oktira Roka Aji, Kenia Permata Sukma, Wiyudi Gomulya</p><br />
<br />
<h3>Visual graphic design team :</h3><br />
<p>Fania Feby Ramadhani</p><br />
<br />
<h3>Wiki :</h3><br />
<p>Muhammad Hariomurti Mardikusumo, Rizky Kusumah</p><br />
<br />
<h3>Human practice team :</h3><br />
<p>Nimas Ghasani, Risma Wiharyanti, Wuddan Nadhirah Rodiana, Bakhtiar Hermawan</p><br />
<br />
<h3>Public Relation :</h3><br />
<p>Mardalisa</p><br />
<br />
<h3>Project Idea :</h3><br />
<p>Tirta Widi Gilang Citradi, Yovin, Christian Heryakusuma</p><br />
<br />
<h3>General Support :</h3><br />
<p>Dr. Sony Suhandono, Dr. Dessy Natalia, Dr. Maelita R. Moeis helped us during our brainstorming<br>Dr. Dessy Natalia is letting us use her lab for some experiments</p><br />
<br />
<h3>Modeling Support :</h3><br />
<p>Melia Silmi and Faisal Rahmananda are bachelor students , department of mathematics help us to model our system<br>Dr. Mochamad Apri is our Instructor and lecturer in department of mathematics. He is an expert on mathematical modelling and helped our team to model our system.</p><br />
<br />
<h3>Imaging Support :</h3><br />
<p>Faculty of Mathematical and Natural Science Institut Teknologi Bandung helped with the Scanning Electron Microscope (SEM) analysis to our degradation system</p><br />
<br />
<h3>Video Support:</h3><br />
<p> Muhammad Daniel Septian, magister student majoring design in Faculty of Arts and Design help us to make a project video</p><br />
<br />
<h3>Wiki Support</h3><br />
<p>Riandy Rahman Nugraha is a bachelor students, department of Informatics Engineering guided us to make our wiki</p> <br />
<br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Modeling
Team:ITB Indonesia/Modeling
2014-10-17T03:12:59Z
<p>Erawijantari: </p>
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia">HOME</a></li><br />
<li>TEAM<br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/FutureSystem">FUTURE SYSTEM</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Degradation Module</h1><br />
<p>The degradation reaction of PET<br>PET+LC Cutinase→Ethylene Glycole +Terephtalic Acid+Cutinase</p><br />
<br />
<ol><br />
<li>Cutinase<br><br />
<p align="justify">The amount of LC-Cutinase enzyme is due to the enzyme production rate by <i>Escherichia coli</i> and the degradation of LC-Cutinase. The enzyme (LC-Cutinase) production occurs constitutively (did not need any activator and produced continuously). At the t-time when LC-Cutinase reach the maximum quantity, the Escherichia coli productivity will be decreased because of Inclusion body. Inclusion body is protein which is produced by miss-folded protein. The amount of inclusion body will be increases. When the inclusion body reach the K quantity, it will be repress the LC-Cutinase production because the inclusion body become a inducer for PibPab promoter that could drive Tet Represor gene expression. Our degradation module contain Tet Operator that could be repressed by Tet Represor. Our PibPab promoter also could drive the Amil Cp gene expression that release blue chromoprotein. Based on that phenomenon, the production of LC-Cutinase was modeled using Hill Function with the repressor factor.</p><br />
<img src="hhttps://static.igem.org/mediawiki/parts/e/e4/ITB-DegModellingJPG.JPG"><br />
<br><br />
</li><br />
<li>Inclusion Body<br><br />
<p align="justify">When the <i>Escherichia coli</i> start to produce many of cutinase enzyme at maximum concentration, it start become stress and begin to producing a miss-folded protein called inclusion body. As long as the producing of enzyme cutinase in this condition, the producing of inclusion body will be much. At this time the inclusion body act as the repressor that stop the production of cutinase and also will be the activator of reporter gen (Amil CP). This activity shown by the picture above. The model expression of inclusion body was modeled using Hill Function.</p><br />
<p>The assumption of the model:<br><br />
We do not include the factor of cell division of bacteria<br>Cutinase</p><br />
<img src="https://static.igem.org/mediawiki/2014/e/e4/Mod2.JPG"><br><br />
<p>Inclusion Body inducer</p><br />
<img src="https://static.igem.org/mediawiki/2014/2/2d/Mod3.JPG"><br><br />
<p>Reporter gen (Amil CP) expression</p><br />
<img src="https://static.igem.org/mediawiki/2014/b/b9/Mod4.JPG"><br><br />
<p>Ethylene Glycole amount (as a product of PET degradation)</p><br />
<img src="https://static.igem.org/mediawiki/2014/2/2f/Mod5.JPG"><br><br />
<p>PET degradation</p><br />
<img src="https://static.igem.org/mediawiki/2014/a/a5/Mod6.JPG"><br><br><br />
<p>Parameters</p><br />
<img src="https://static.igem.org/mediawiki/2014/a/a5/Mod6.JPG"><br><br><br />
<p align="justify">Figure 1 shown the graph of cutinase production, inclussion body production and blue protein expression. Figure 2 shown the the close up graph and Figure 3 shown the PET degradation and ethylene glicol product profil by the time of gene expression. Inclussion bodies are reflactile aggreates of protease-resistant misfolded protein that often occur in recombinant bacteria upon gratutious overexpression of cloned genes. In biotechnology, the formation of Ibs represents a main obstacle for protein production since even favouring high protein yields. Aggregation of recombinant proteins is probably due to limiting amount of chaperones when recombinant gene expression is directed at high leves. When the ammount of protein production is maximal, we assume that inclusion body will be release. Inclussion body could drive the expression of Tet represor and AmilCp as shown in figure 1. When the inclussion body could be degraded by the cell, the PibPab promoter will be off and there are no TetR and amilCP expression. In that condition we assume that the the cutinase expression will be increassing until reach some value that could release the inclusion body, than the expression will be represed. In figure 2, we could observe the osilation graph. From the figure 3, we could see that by the time of PET degradation the ethylene glicol product that could be released is increase.</p><br><br />
<img src="https://static.igem.org/mediawiki/2014/6/66/Mod9.JPG"><br><br />
<img src="https://static.igem.org/mediawiki/2014/5/5c/Mod10.JPG"><br><br />
</li><br />
<p<br />
<br><br />
<br />
<li>References<br />
<ol><br />
<li>Carrio, M.M and A. Villaverde. 2002. Construction and Deconstruction of Bacterial Inclusion Bodies. <i>Journal of Biotechnology 96. 3-12</i></li><br />
<li>Chapman, and Halu/CRC, “AN INTRODUCTION TO SYSTEMS BIOLOGY Design Principle of Biological Circuit”</li><br />
<li>Fink, Anthony L. 1998. Protein Aggregation: folding aggregates, inclusion bodies and amyloid. <i>© Current Biology Ltd ISSN 1359-0278</i></li><br />
</ol><br />
</li><br />
</ol><br />
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Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Data
Team:ITB Indonesia/Data
2014-10-17T03:11:46Z
<p>Erawijantari: </p>
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<h1>A. Scaning Electron Microscope (SEM) Result Analysis</h1><br />
<p align="justify">For biodegrading experiments, we used 3x5 cm2 bottle plastics with average weight 0.05 g. Then, we washed it several times with water and ethanol.</p><br />
<p align="justify">Plastic sample was applied just before we inoculated the bacteria on the medium. Bacterial culture were grown at 37C in Luria-Broth medium. After 3 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss.</p><br />
<p align="justify">Meanwhile, for UC Davis bacterial culture, we inoculated the bacteria in Luria-Broth medium supplemented with 170 ppm cloramphenicol. After an absorbance A600 nm of 0.6 was reached, we added 1% of arabinose and plastic sample. After 2 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss.</p><br />
<p align="justify">We used medium supplemented with antibiotics and plastic sample as control on this experiment. The scaning electron microsccopy analysis of fractured surface of PET film was carried out using Scaning electron microscope. The surface of the treated PET samples were coated with conductive heavy metals such as gold/palladium.</p><br />
<palign="justify">SEM image shows that cracks were observed at the surface of a plastic sample (PET) after incubation on the bacterial culture. But, there is no significant weight decreased of the plastics samples. From this SEM study we conclude that both our LC Cut UC Davis and OmpA-LC-Cut Fussion ITB 2014 able to degrade PET plastics samples. Figure 1. shown the control sampel that we could not observe any cracks. Figure 2 and 3 shown the PET degradation by Lc cutinase from UC davis and from ITB_Indonesia.</p><br />
<br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/e/e9/Sem1.JPG"><br />
<p><small>Figure 1. Control sampel (PET plastic incubated in medium)</small></p><br />
</div><br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/9/92/Sem2.JPG"><br />
<p><small>Figure 2. PET plastic after treatment with ompA-LC-cutinase from ITB_Indonesia construct (BBa_K1387006)</small></p><br />
</div><br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/6/62/Sem3.JPG"><br />
<p><small>Figure 3. PET plastic after treatment with LC-cutinase UC Davis 2012 gene construct (Bba_K936020)</small></p><br />
</div><br />
<br />
<h3>References</h3><br />
<ol style="text-decoration:none;"><br />
<li>Sepperumal, Umaheswari, Murali Markandan, and Anbusaravanan Natarajan. 2013. electron microscopic studies of Polyethylene terepthalate degradation potential of Pseudomonas species. J. Microbiol. Biotech. Res. (1): 104-110</li><br />
<li>Mittal,Alok, R.K. Soni, Khrisna Dutt, and Swati Singh. 2010. Scaning electron microscopy study of hazardous waste flakes of polyethylene terephthalate (PET) by aminolysis and ammonolysis. Journal of Hazardous Materials Volume 178, Issues 1-3 </li><br />
</ol><br />
<br />
<br><br />
<h1>B. Ethylene Glikol Chromic Acid Assay</h1><br />
<p align="justify">Poly(ethylene terephthalate) (PET) is a plastic that is a polymer of ethylene terephthalate (C10H8O4) units (Sarker et al., 2010). Polyethylene terephthalate (PET) can be degrade via limited enzymatic hydrolysis of the ester bond of the polymer backbone, one enzyme that have been assesed for this purpose is cutinase (Ribitsch et al., 2012). When polyethylene terephthalate degraded, it will produce terephthalic acid and ethylene glycol (Venkatachalam et al., 2012). Ethylene glycol is one of alcohol compound. Oxidation of alcohols by strong oxidants such as chromic acid (K2Cr2O7) in suphuric acid(H2SO4) is possible, but differs depending on the degree of alcohol. If a reaction has occurred using chromic acid in sulphuric acid, there is a color change from orange to green. In our project we use chromic acid to detect ethylene glycol as product of the PET degradation by LC cutinase enzyme.</p><br />
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<p align="justify">Each sample used for measure the absorbance contain a fixed amount of chromic acid. The absorbance of the chromic acid will represent the total amount of ethylene glycol inversely. It means that if the absorbance is high, the amount of ethylene glycol in solution is low, and if the absorbance is low, the amount of ethylene glycol in solution is high. If the absorbance is high, the amount of chromic acid is high, which means that the amount of ethylene glycol being oxidized by chromic acid is low and vice versa. Both of LC Cutinase UC Davis and OmpA-LC Cutinase ITB_Indonesia shown the degrading activity of PET become ethylene glycol as once of the product.The optimum time to produce ethylene glycol is on the fourth hour after inoculation.</p><br />
<br />
<h3>References</h3><br />
<ol style="text-decoration:none;"><br />
<li>Ribitsch D, Enrique HA, Katrin G, Anita D, Sabine Z, Annemarie M, Rosario DR, Georg S, Karl G, Helmut S and Georg MG. 2012. A New Esterase from Thermobifida halotolerans Hydrolyses Polyethylene Terephthalate (PET) and Polylactic Acid (PLA). <i>Polymers. 4: 617-629</i></li><br />
<li>Sarker M, Aminul K, Mohammad MR, Mohammed M and ASMD Mohammad. 2011. Waste Polyethylene Terephthalate (PETE-1) Conversion into Liquid Fuel. <i>Journal of Fundamentals of Renewable Energy and Applications. 1</i></li><br />
<li>Venkatachalam S, Shilpa GN, Jayprakash VL, Prashant RG, Krishna R and Anil KK. 2012. Degradation and Recyclability of Poly (Ethylene Terephthalate). <i>Intech</i></li><br />
</ol><br />
<br />
<br><br />
<h1>C. pNPP assay</h1><br />
<p align="justify">Blastp program shows that LC cutinase shows high amino acid sequence similarity (54 to 60%) to lipase (Sulaiman, 2012). So, we tried to measure LC-cutinase activity using p-nitrophenylpalmitate (pNPP) as a substrate. pNPP typically used for measuring lipase activity.</p><br />
<p>In this assay, we used bacterial culture because in real application we would like to use bacterial culture to degrade PETdirectly. As negative control, we used <i>E.coli </i> BL21(DE3) without plasmid. We also included LC-cutinase UC Davis 2012 gene construct (Bba_K936020) in this experiment. LC-cutinase UC Davis 2012 bacterial culture is induced after 2 hours of growth (after OD 600 nm ±0.6 was reached) using 1% of arabinose. All of the bacterial culture were harvested after 4 hours of growth.From ethylene glycol assay using chromic acid, we found that ompA-LC-cutinase from ITB_Indonesia construct (BBa_K1387006) has the highest activity after 4 hours of growth.</p><br />
<p align="justify">At first, we measured the activity at 60 degrees Celsius. We found that both ompA-LC-cutinase from ITB_Indonesia and LC-cutinase UC Davis 2012 still performed small activity. Then, we tried measure the activity at 25 degrees Celsius. The ompA-LC-cutinase from ITB_Indonesia’s activity was 0.001 U/mL. While, LC-cutinase UC Davis 2012 has ten fold higher (0.01 U/mL). The LC cutinase activity was determined based on the standard curve of p-nitrophenol. One unit of cutinase activity was defined as the amount of enzyme releasing 1 μmol pNP per minute under the assay conditions.</p><br />
<p align="justify">From this experiment, we are able to confirm that ompA-LC-cutinase from ITB_Indonesia successfully perform activity. But, its suggest that we still have to check the possible activity in different time of bacterial growth, expression rate, LC-cutinase presentation by ompA and activity assay using different substrate.</p><br />
<br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/9/90/Sem6.JPG"><br />
<p><small>Figure 1. LC Cutinase activity. The enzymatic activity was determined at 60C in assay condition using pNP-palmitate (C16) as a substrate. The experiment was carried out twice.</small></p><br />
</div><br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/2014/a/a1/Sem7.JPG"><br />
<p><small>Figure 2. LC Cutinase activity. The enzymatic activity was determined at 25 C in assay condition using pNP-palmitate (C16) as a substrate. The experiment was carried out twice.</small></p><br />
</div><br />
<p align"justify">Small activity of LC-Cutinase maybe due to their oligomeric state. Tethering protein to cell surface may affect folding of protein and also their biological function (Park,2010). These suggest us to look futher detail on folding and displaying state of LC cutinase.</p><br />
<br />
<p align="justify">There are several things that should be noted when we designate fusion strategy of passanger protein or protein of interest with the outer membrane of protein. Linking the different passanger proteins to the same outer membrane protein may result in different translocation. The characteristic of passanger protein affects the translocation. Its characteristic such as formation of disulfide bridge, total residue of hydrophobic aminoacid etc. </p><br />
<p align="justify">So there are four requirements have to be met for constructing well designed surface display. First is, it has to possesses efficient signal peptide, 2nd it should have strong anchor to keep the fusion protein from detachment. 3rd it should be compatible to the sequence that is inserted or fused. The last is it should resisstant to the attack of the protease from medium or periplasmic membrane. </p><br />
<br />
<h3>Reference</h3><br />
<ol style="text-decoration:none;"><br />
<li>Sulaiman, Sintawee, SayaYamat, EikoKanaya, Joong-Jae Kim, Yuichi Koga, Kazufumi Takano and Shigenori Kanaya. 2012. Isolation of a Novel Cutinase Homolog with Polyethylene Terephthalate-Degrading Activity from Leaf-Branch Compost by Using a Metagenomic Approach. <i>Applied and Environmental Microbiology p 1556-1562.</i></li><br />
<li>Lee, Sang Yup., Choi, Jong Hyun., Xu, Zhaohui. 2003. “Microbial Cell-Surface Display” <i>Elsevier</i></li><br />
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<br />
<br><br />
<h1>Data</h1><br />
<ol type="A"><br />
<h3><br><li>Ethylene Glycol Assay using Chromic Acid</h3></li><br />
<p>Poly(ethylene terephthalate) (PET) is a plastic that is a polymer of ethylene terephthalate (C10H8O4) units (Sarker et al., 2010). Polyethylene terephthalate (PET) can be degrade via limited enzymatic hydrolysis of the ester bond of the polymer backbone, one enzyme that have been assesed for this purpose is cutinase (Ribitsch et al., 2012). When polyethylene terephthalate degraded, it will produce terephthalic acid and ethylene glycol (Venkatachalam et al., 2012). Ethylene glycol is one of alcohol compound. Oxidation of alcohols by strong oxidants such as chromic acid (K2Cr2O7) in suphuric acid(H2SO4) is possible, but differs depending on the degree of alcohol. If a reaction has occurred using chromic acid in sulphuric acid, there is a color change from orange to green. In our project we use chromic acid to detect ethylene glycol as product of the PET degradation by LC cutinase enzyme.figure 1 shown the result of ethylene glycol assay using chromic acid.</p><br />
<p><img src="https://static.igem.org/mediawiki/parts/7/79/Rumus.JPG" width="300"/><p><br />
<div style="text-align: center;"><br />
<p><img src="https://static.igem.org/mediawiki/parts/f/fb/Graph.JPG" width="500"/></p><br />
<p><small>Figure 1. Ethylene Glycol Concentration LC ITB and LC UC Davis<br />
</small></p><br />
</div><br />
<p>Each sample used for measure the absorbance contain a fixed amount of chromic acid. The absorbance of the chromic acid will represent the total amount of ethylene glycol inversely. It means that if the absorbance is high, the amount of ethylene glycol in solution is low, and if the absorbance is low, the amount of ethylene glycol in solution is high. If the absorbance is high, the amount of chromic acid is high, which means that the amount of ethylene glycol being oxidized by chromic acid is low and vice versa. Both of LC Cutinase UC Davis and OmpA-LC Cutinase ITB_Indonesia shown the degrading activity of PET become ethylene glycol as once of the product.The optimum time to produce ethylene glycol is on the fourth hour after inoculation. </p><br />
<p> Reference</p><br />
<p>Ribitsch D, Enrique HA, Katrin G, Anita D, Sabine Z, Annemarie M, Rosario DR, Georg S, Karl G, Helmut S and Georg MG. 2012. A New Esterase from Thermobifida halotolerans Hydrolyses Polyethylene Terephthalate (PET) and Polylactic Acid (PLA). Polymers. 4: 617-629 </p><br />
<p>Sarker M, Aminul K, Mohammad MR, Mohammed M and ASMD Mohammad. 2011. Waste Polyethylene Terephthalate (PETE-1) Conversion into Liquid Fuel. Journal of Fundamentals of Renewable Energy and Applications. 1</p><br />
<p>Venkatachalam S, Shilpa GN, Jayprakash VL, Prashant RG, Krishna R and Anil KK. 2012. Degradation and Recyclability of Poly (Ethylene Terephthalate). Intech</p><br />
<p><h3><br><li> Scaning Electron Microscope (SEM) Result Analysis </h3></li></p><br />
<p>For biodegrading experiments, we used 3x5 cm2 bottle plastics with average weight 0.05 g. Then, we washed it several times with water and ethanol. </p><br />
<p>Plastic sample was applying just before we inoculated the bacteria on the medium. Bacterial culture were grown at 37C in Luria-Broth medium. After 3 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss. </p><br />
<p>Meanwhile, for UC Davis bacterial culture, we inoculated the bacteria in Luria-Broth medium supplemented with 170 ppm cloramphenicol. After an absorbance A600 nm of 0.6 was reached, we added 1% of arabinose and plastic sample. After 2 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss.</p><br />
<p>We used medium supplemented with antibiotics and plastic sample as control on this experiment. The scaning electron microsccopy analysis of fractured surface of PET film was carried out using Scaning electron microscope. The surface of the treated PET samples were coated with conductive heavy metals such as gold/palladium. </p><br />
<p>SEM image shows that cracks were observed at the surface of a plastic sample (PET) after incubation on the bacterial culture. But, there is no significant weight decreased of the plastics samples. From this SEM study we conclude that both our LC Cut UC Davis and OmpA-LC-Cut Fussion ITB 2014 able to degrade PET plastics samples. Figure 1. shown the control sampel that we could not observe any cracks. Figure 2 and 3 shown the PET degradation by Lc cutinase from UC davis and from ITB_Indonesia. </p><br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/parts/4/4e/SEMcontrol.jpg" width="500"/><br />
<p><small>Figure 1. Control sample (E.coli BL21 without plasmid)</small></p><br />
</div><br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/parts/2/2e/SEMUCDavis.jpg" width="500"/><br />
<p><small>Figure 2. PET plastic after treatment with LC-cutinase UC Davis 2012 gene construct (Bba_K936020) </small></p><br />
</div> <br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/parts/c/c1/SEMITBIndonesia.jpg" width="500"/><br />
<p><small>Figure 3. PET plastic after treatment with ompA-LC-cutinase from ITB_Indonesia construct (BBa_K1387006) </small></p><br />
</div> <br />
<p>Reference</p><br />
<p>Sepperumal, Umaheswari, Murali Markandan, and Anbusaravanan Natarajan. 2013. electron microscopic studies of Polyethylene terepthalate degradation potential of Pseudomonas species. J. Microbiol. Biotech. Res. (1): 104-110</p><br />
<p>Mittal,Alok, R.K. Soni, Khrisna Dutt, and Swati Singh. 2010. Scaning electron microscopy study of hazardous waste flakes of polyethylene terephthalate (PET) by aminolysis and ammonolysis. Journal of Hazardous Materials Volume 178, Issues 1-3 </p><br />
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Erawijantari
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Team:ITB Indonesia/Data
2014-10-15T16:18:02Z
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<br />
<br><br />
<h1>Data</h1><br />
<ol type="A"><br />
<h3><br><li>Ethylene Glycol Assay using Chromic Acid</h3></li><br />
<p>Poly(ethylene terephthalate) (PET) is a plastic that is a polymer of ethylene terephthalate (C10H8O4) units (Sarker et al., 2010). Polyethylene terephthalate (PET) can be degrade via limited enzymatic hydrolysis of the ester bond of the polymer backbone, one enzyme that have been assesed for this purpose is cutinase (Ribitsch et al., 2012). When polyethylene terephthalate degraded, it will produce terephthalic acid and ethylene glycol (Venkatachalam et al., 2012). Ethylene glycol is one of alcohol compound. Oxidation of alcohols by strong oxidants such as chromic acid (K2Cr2O7) in suphuric acid(H2SO4) is possible, but differs depending on the degree of alcohol. If a reaction has occurred using chromic acid in sulphuric acid, there is a color change from orange to green. In our project we use chromic acid to detect ethylene glycol as product of the PET degradation by LC cutinase enzyme.figure 1 shown the result of ethylene glycol assay using chromic acid.</p><br />
<p><img src="https://static.igem.org/mediawiki/parts/7/79/Rumus.JPG" width="300"/><p><br />
<div style="text-align: center;"><br />
<p><img src="https://static.igem.org/mediawiki/parts/f/fb/Graph.JPG" width="500"/></p><br />
<p><small>Figure 1. Ethylene Glycol Concentration LC ITB and LC UC Davis<br />
</small></p><br />
</div><br />
<p>Each sample used for measure the absorbance contain a fixed amount of chromic acid. The absorbance of the chromic acid will represent the total amount of ethylene glycol inversely. It means that if the absorbance is high, the amount of ethylene glycol in solution is low, and if the absorbance is low, the amount of ethylene glycol in solution is high. If the absorbance is high, the amount of chromic acid is high, which means that the amount of ethylene glycol being oxidized by chromic acid is low and vice versa. Both of LC Cutinase UC Davis and OmpA-LC Cutinase ITB_Indonesia shown the degrading activity of PET become ethylene glycol as once of the product.The optimum time to produce ethylene glycol is on the fourth hour after inoculation. </p><br />
<p> Reference</p><br />
<p>Ribitsch D, Enrique HA, Katrin G, Anita D, Sabine Z, Annemarie M, Rosario DR, Georg S, Karl G, Helmut S and Georg MG. 2012. A New Esterase from Thermobifida halotolerans Hydrolyses Polyethylene Terephthalate (PET) and Polylactic Acid (PLA). Polymers. 4: 617-629 </p><br />
<p>Sarker M, Aminul K, Mohammad MR, Mohammed M and ASMD Mohammad. 2011. Waste Polyethylene Terephthalate (PETE-1) Conversion into Liquid Fuel. Journal of Fundamentals of Renewable Energy and Applications. 1</p><br />
<p>Venkatachalam S, Shilpa GN, Jayprakash VL, Prashant RG, Krishna R and Anil KK. 2012. Degradation and Recyclability of Poly (Ethylene Terephthalate). Intech</p><br />
<p><h3><br><li> Scaning Electron Microscope (SEM) Result Analysis </h3></li></p><br />
<p>For biodegrading experiments, we used 3x5 cm2 bottle plastics with average weight 0.05 g. Then, we washed it several times with water and ethanol. </p><br />
<p>Plastic sample was applying just before we inoculated the bacteria on the medium. Bacterial culture were grown at 37C in Luria-Broth medium. After 3 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss. </p><br />
<p>Meanwhile, for UC Davis bacterial culture, we inoculated the bacteria in Luria-Broth medium supplemented with 170 ppm cloramphenicol. After an absorbance A600 nm of 0.6 was reached, we added 1% of arabinose and plastic sample. After 2 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss.</p><br />
<p>We used medium supplemented with antibiotics and plastic sample as control on this experiment. The scaning electron microsccopy analysis of fractured surface of PET film was carried out using Scaning electron microscope. The surface of the treated PET samples were coated with conductive heavy metals such as gold/palladium. </p><br />
<p>SEM image shows that cracks were observed at the surface of a plastic sample (PET) after incubation on the bacterial culture. But, there is no significant weight decreased of the plastics samples. From this SEM study we conclude that both our LC Cut UC Davis and OmpA-LC-Cut Fussion ITB 2014 able to degrade PET plastics samples. Figure 1. shown the control sampel that we could not observe any cracks. Figure 2 and 3 shown the PET degradation by Lc cutinase from UC davis and from ITB_Indonesia. </p><br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/parts/4/4e/SEMcontrol.jpg" width="500"/><br />
<p><small>Figure 1. Control sample (E.coli BL21 without plasmid)</small></p><br />
</div><br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/parts/2/2e/SEMUCDavis.jpg" width="500"/><br />
<p><small>Figure 2. PET plastic after treatment with LC-cutinase UC Davis 2012 gene construct (Bba_K936020) </small></p><br />
</div> <br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/parts/c/c1/SEMITBIndonesia.jpg" width="500"/><br />
<p><small>Figure 3. PET plastic after treatment with ompA-LC-cutinase from ITB_Indonesia construct (BBa_K1387006) </small></p><br />
</div> <br />
<br />
<br />
<br></div>
Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Data
Team:ITB Indonesia/Data
2014-10-15T16:16:37Z
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
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<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Data</h1><br />
<ol type="A"><br />
<h3><br><li>Ethylene Glycol Assay using Chromic Acid</h3></li><br />
<p>Poly(ethylene terephthalate) (PET) is a plastic that is a polymer of ethylene terephthalate (C10H8O4) units (Sarker et al., 2010). Polyethylene terephthalate (PET) can be degrade via limited enzymatic hydrolysis of the ester bond of the polymer backbone, one enzyme that have been assesed for this purpose is cutinase (Ribitsch et al., 2012). When polyethylene terephthalate degraded, it will produce terephthalic acid and ethylene glycol (Venkatachalam et al., 2012). Ethylene glycol is one of alcohol compound. Oxidation of alcohols by strong oxidants such as chromic acid (K2Cr2O7) in suphuric acid(H2SO4) is possible, but differs depending on the degree of alcohol. If a reaction has occurred using chromic acid in sulphuric acid, there is a color change from orange to green. In our project we use chromic acid to detect ethylene glycol as product of the PET degradation by LC cutinase enzyme.figure 1 shown the result of ethylene glycol assay using chromic acid.</p><br />
<p><img src="https://static.igem.org/mediawiki/parts/7/79/Rumus.JPG" width="300"/><p><br />
<div style="text-align: center;"><br />
<p><img src="https://static.igem.org/mediawiki/parts/f/fb/Graph.JPG" width="500"/></p><br />
<p><small>Figure 1. Ethylene Glycol Concentration LC ITB and LC UC Davis<br />
</small></p><br />
</div><br />
<p>Each sample used for measure the absorbance contain a fixed amount of chromic acid. The absorbance of the chromic acid will represent the total amount of ethylene glycol inversely. It means that if the absorbance is high, the amount of ethylene glycol in solution is low, and if the absorbance is low, the amount of ethylene glycol in solution is high. If the absorbance is high, the amount of chromic acid is high, which means that the amount of ethylene glycol being oxidized by chromic acid is low and vice versa. Both of LC Cutinase UC Davis and OmpA-LC Cutinase ITB_Indonesia shown the degrading activity of PET become ethylene glycol as once of the product.The optimum time to produce ethylene glycol is on the fourth hour after inoculation. </p><br />
<p> Reference</p><br />
<p>Ribitsch D, Enrique HA, Katrin G, Anita D, Sabine Z, Annemarie M, Rosario DR, Georg S, Karl G, Helmut S and Georg MG. 2012. A New Esterase from Thermobifida halotolerans Hydrolyses Polyethylene Terephthalate (PET) and Polylactic Acid (PLA). Polymers. 4: 617-629 </p><br />
<p>Sarker M, Aminul K, Mohammad MR, Mohammed M and ASMD Mohammad. 2011. Waste Polyethylene Terephthalate (PETE-1) Conversion into Liquid Fuel. Journal of Fundamentals of Renewable Energy and Applications. 1</p><br />
<p>Venkatachalam S, Shilpa GN, Jayprakash VL, Prashant RG, Krishna R and Anil KK. 2012. Degradation and Recyclability of Poly (Ethylene Terephthalate). Intech</p><br />
<p><h3><br><li> Scaning Electron Microscope (SEM) Result Analysis </h3></li></p><br />
<p>For biodegrading experiments, we used 3x5 cm2 bottle plastics with average weight 0.05 g. Then, we washed it several times with water and ethanol. </p><br />
<p>Plastic sample was applying just before we inoculated the bacteria on the medium. Bacterial culture were grown at 37C in Luria-Broth medium. After 3 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss. </p><br />
<p>Meanwhile, for UC Davis bacterial culture, we inoculated the bacteria in Luria-Broth medium supplemented with 170 ppm cloramphenicol. After an absorbance A600 nm of 0.6 was reached, we added 1% of arabinose and plastic sample. After 2 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss.</p><br />
<p>We used medium supplemented with antibiotics and plastic sample as control on this experiment. The scaning electron microsccopy analysis of fractured surface of PET film was carried out using Scaning electron microscope. The surface of the treated PET samples were coated with conductive heavy metals such as gold/palladium. </p><br />
<p>SEM image shows that cracks were observed at the surface of a plastic sample (PET) after incubation on the bacterial culture. But, there is no significant weight decreased of the plastics samples. From this SEM study we conclude that both our LC Cut UC Davis and OmpA-LC-Cut Fussion ITB 2014 able to degrade PET plastics samples. Figure 1. shown the control sampel that we could not observe any cracks. Figure 2 and 3 shown the PET degradation by Lc cutinase from UC davis and from ITB_Indonesia. </p><br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/parts/4/4e/SEMcontrol.jpg" width="500"/><br />
<p><small>Figure 1. Control sample (E.coli BL21 without plasmid) (</small></p><br />
</div><br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/parts/2/2e/SEMUCDavis.jpg" width="500"/><br />
<p><small>Figure 2. PET plastic after treatment with LC-cutinase UC Davis 2012 gene construct (Bba_K936020) (</small></p><br />
</div> <br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/parts/c/c1/SEMITBIndonesia.jpg" width="500"/><br />
<p><small>Figure 3. PET plastic after treatment with ompA-LC-cutinase from ITB_Indonesia construct (BBa_K1387006) (</small></p><br />
</div> <br />
<br />
<br />
<br></div>
Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Data
Team:ITB Indonesia/Data
2014-10-15T16:11:08Z
<p>Erawijantari: </p>
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</div><br />
<br />
<br><br />
<h1>Data</h1><br />
<ol type="A"><br />
<h3><br><li>Ethylene Glycol Assay using Chromic Acid</h3></li><br />
<p>Poly(ethylene terephthalate) (PET) is a plastic that is a polymer of ethylene terephthalate (C10H8O4) units (Sarker et al., 2010). Polyethylene terephthalate (PET) can be degrade via limited enzymatic hydrolysis of the ester bond of the polymer backbone, one enzyme that have been assesed for this purpose is cutinase (Ribitsch et al., 2012). When polyethylene terephthalate degraded, it will produce terephthalic acid and ethylene glycol (Venkatachalam et al., 2012). Ethylene glycol is one of alcohol compound. Oxidation of alcohols by strong oxidants such as chromic acid (K2Cr2O7) in suphuric acid(H2SO4) is possible, but differs depending on the degree of alcohol. If a reaction has occurred using chromic acid in sulphuric acid, there is a color change from orange to green. In our project we use chromic acid to detect ethylene glycol as product of the PET degradation by LC cutinase enzyme.figure 1 shown the result of ethylene glycol assay using chromic acid.</p><br />
<p><img src="https://static.igem.org/mediawiki/parts/7/79/Rumus.JPG" width="300"/><p><br />
<div style="text-align: center;"><br />
<p><img src="https://static.igem.org/mediawiki/parts/f/fb/Graph.JPG" width="500"/></p><br />
<p><small>Figure 1. Ethylene Glycol Concentration LC ITB and LC UC Davis<br />
</small></p><br />
</div><br />
<p>Each sample used for measure the absorbance contain a fixed amount of chromic acid. The absorbance of the chromic acid will represent the total amount of ethylene glycol inversely. It means that if the absorbance is high, the amount of ethylene glycol in solution is low, and if the absorbance is low, the amount of ethylene glycol in solution is high. If the absorbance is high, the amount of chromic acid is high, which means that the amount of ethylene glycol being oxidized by chromic acid is low and vice versa. Both of LC Cutinase UC Davis and OmpA-LC Cutinase ITB_Indonesia shown the degrading activity of PET become ethylene glycol as once of the product.The optimum time to produce ethylene glycol is on the fourth hour after inoculation. </p><br />
<p> Reference</p><br />
<p>Ribitsch D, Enrique HA, Katrin G, Anita D, Sabine Z, Annemarie M, Rosario DR, Georg S, Karl G, Helmut S and Georg MG. 2012. A New Esterase from Thermobifida halotolerans Hydrolyses Polyethylene Terephthalate (PET) and Polylactic Acid (PLA). Polymers. 4: 617-629 </p><br />
<p>Sarker M, Aminul K, Mohammad MR, Mohammed M and ASMD Mohammad. 2011. Waste Polyethylene Terephthalate (PETE-1) Conversion into Liquid Fuel. Journal of Fundamentals of Renewable Energy and Applications. 1</p><br />
<p>Venkatachalam S, Shilpa GN, Jayprakash VL, Prashant RG, Krishna R and Anil KK. 2012. Degradation and Recyclability of Poly (Ethylene Terephthalate). Intech</p><br />
<p><h3><br><li> Scaning Electron Microscope (SEM) Result Analysis </h3></li></p><br />
<p>For biodegrading experiments, we used 3x5 cm2 bottle plastics with average weight 0.05 g. Then, we washed it several times with water and ethanol. </p><br />
<p>Plastic sample was applying just before we inoculated the bacteria on the medium. Bacterial culture were grown at 37C in Luria-Broth medium. After 3 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss. </p><br />
<p>Meanwhile, for UC Davis bacterial culture, we inoculated the bacteria in Luria-Broth medium supplemented with 170 ppm cloramphenicol. After an absorbance A600 nm of 0.6 was reached, we added 1% of arabinose and plastic sample. After 2 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss.</p><br />
<p>We used medium supplemented with antibiotics and plastic sample as control on this experiment. The scaning electron microsccopy analysis of fractured surface of PET film was carried out using Scaning electron microscope. The surface of the treated PET samples were coated with conductive heavy metals such as gold/palladium. </p><br />
<p>SEM image shows that cracks were observed at the surface of a plastic sample (PET) after incubation on the bacterial culture. But, there is no significant weight decreased of the plastics samples. From this SEM study we conclude that both our LC Cut UC Davis and OmpA-LC-Cut Fussion ITB 2014 able to degrade PET plastics samples. Figure 1. shown the control sampel that we could not observe any cracks. Figure 2 and 3 shown the PET degradation by Lc cutinase from UC davis and from ITB_Indonesia. </p><br />
<div style="text-align: center;"><br />
<img src="https://static.igem.org/mediawiki/parts/4/4e/SEMcontrol.jpg" width="500"/><br />
<p><small>Figure 1. Control sample (E.coli BL21 without plasmid) (</small></p><br />
</div><br />
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<h1>Data</h1><br />
<ol type="A"><br />
<h3><br><li>Ethylene Glycol Assay using Chromic Acid</h3></li><br />
<p>Poly(ethylene terephthalate) (PET) is a plastic that is a polymer of ethylene terephthalate (C10H8O4) units (Sarker et al., 2010). Polyethylene terephthalate (PET) can be degrade via limited enzymatic hydrolysis of the ester bond of the polymer backbone, one enzyme that have been assesed for this purpose is cutinase (Ribitsch et al., 2012). When polyethylene terephthalate degraded, it will produce terephthalic acid and ethylene glycol (Venkatachalam et al., 2012). Ethylene glycol is one of alcohol compound. Oxidation of alcohols by strong oxidants such as chromic acid (K2Cr2O7) in suphuric acid(H2SO4) is possible, but differs depending on the degree of alcohol. If a reaction has occurred using chromic acid in sulphuric acid, there is a color change from orange to green. In our project we use chromic acid to detect ethylene glycol as product of the PET degradation by LC cutinase enzyme.figure 1 shown the result of ethylene glycol assay using chromic acid.</p><br />
<p><img src="https://static.igem.org/mediawiki/parts/7/79/Rumus.JPG" width="300"/><p><br />
<div style="text-align: center;"><br />
<p><img src="https://static.igem.org/mediawiki/parts/f/fb/Graph.JPG" width="500"/></p><br />
<p><small>Figure 1. Ethylene Glycol Concentration LC ITB and LC UC Davis<br />
</small></p><br />
</div><br />
<p>Each sample used for measure the absorbance contain a fixed amount of chromic acid. The absorbance of the chromic acid will represent the total amount of ethylene glycol inversely. It means that if the absorbance is high, the amount of ethylene glycol in solution is low, and if the absorbance is low, the amount of ethylene glycol in solution is high. If the absorbance is high, the amount of chromic acid is high, which means that the amount of ethylene glycol being oxidized by chromic acid is low and vice versa. Both of LC Cutinase UC Davis and OmpA-LC Cutinase ITB_Indonesia shown the degrading activity of PET become ethylene glycol as once of the product.The optimum time to produce ethylene glycol is on the fourth hour after inoculation. </p><br />
<p> Reference</p><br />
<p>Ribitsch D, Enrique HA, Katrin G, Anita D, Sabine Z, Annemarie M, Rosario DR, Georg S, Karl G, Helmut S and Georg MG. 2012. A New Esterase from Thermobifida halotolerans Hydrolyses Polyethylene Terephthalate (PET) and Polylactic Acid (PLA). Polymers. 4: 617-629 </p><br />
<p>Sarker M, Aminul K, Mohammad MR, Mohammed M and ASMD Mohammad. 2011. Waste Polyethylene Terephthalate (PETE-1) Conversion into Liquid Fuel. Journal of Fundamentals of Renewable Energy and Applications. 1</p><br />
<p>Venkatachalam S, Shilpa GN, Jayprakash VL, Prashant RG, Krishna R and Anil KK. 2012. Degradation and Recyclability of Poly (Ethylene Terephthalate). Intech</p><br />
<p><h3><br><li> Scaning Electron Microscope (SEM) Result Analysis </h3></li></p><br />
<p>For biodegrading experiments, we used 3x5 cm2 bottle plastics with average weight 0.05 g. Then, we washed it several times with water and ethanol. </p><br />
<p>Plastic sample was applying just before we inoculated the bacteria on the medium. Bacterial culture were grown at 37C in Luria-Broth medium. After 3 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss. </p><br />
<p>Meanwhile, for UC Davis bacterial culture, we inoculated the bacteria in Luria-Broth medium supplemented with 170 ppm cloramphenicol. After an absorbance A600 nm of 0.6 was reached, we added 1% of arabinose and plastic sample. After 2 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss.</p><br />
<p>We used medium supplemented with antibiotics and plastic sample as control on this experiment. The scaning electron microsccopy analysis of fractured surface of PET film was carried out using Scaning electron microscope. The surface of the treated PET samples were coated with conductive heavy metals such as gold/palladium. </p><br />
<p>SEM image shows that cracks were observed at the surface of a plastic sample (PET) after incubation on the bacterial culture. But, there is no significant weight decreased of the plastics samples. From this SEM study we conclude that both our LC Cut UC Davis and OmpA-LC-Cut Fussion ITB 2014 able to degrade PET plastics samples. Figure 1. shown the control sampel that we could not observe any cracks. Figure 2 and 3 shown the PET degradation by Lc cutinase from UC davis and from ITB_Indonesia. </p><br />
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Erawijantari
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Team:ITB Indonesia/Data
2014-10-15T15:57:48Z
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<li><a href="https://igem.org/Team.cgi?id=1387" >OFFICIAL TEAM PROFILE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Team">TEAM MEMBER</a></li><br />
</ul><br />
</li><br />
<li>PROJECT<br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Project">OVERVIEW</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/DegMod">DEGRADATION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ConMod">CONVERTION MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/RepMod">REPORTER MODULE</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Parts">PARTS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Attributions">ATTRIBUTIONS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Safety">SAFETY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
</li><br />
<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-modeling">MODELING</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
</ul><br />
</li><br />
<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Medsos">SOCIAL MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/OnMedia">MEDIA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Biosafety">BIOSAFETY SEMINAR</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/MeetUp">INDONESIA TEAMS MEET UP</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Data</h1><br />
<ol type="A"><br />
<h3><br><li>Ethylene Glycol Assay using Chromic Acid</h3></li><br />
<p>Poly(ethylene terephthalate) (PET) is a plastic that is a polymer of ethylene terephthalate (C10H8O4) units (Sarker et al., 2010). Polyethylene terephthalate (PET) can be degrade via limited enzymatic hydrolysis of the ester bond of the polymer backbone, one enzyme that have been assesed for this purpose is cutinase (Ribitsch et al., 2012). When polyethylene terephthalate degraded, it will produce terephthalic acid and ethylene glycol (Venkatachalam et al., 2012). Ethylene glycol is one of alcohol compound. Oxidation of alcohols by strong oxidants such as chromic acid (K2Cr2O7) in suphuric acid(H2SO4) is possible, but differs depending on the degree of alcohol. If a reaction has occurred using chromic acid in sulphuric acid, there is a color change from orange to green. In our project we use chromic acid to detect ethylene glycol as product of the PET degradation by LC cutinase enzyme.figure 1 shown the result of ethylene glycol assay using chromic acid.</p><br />
<p><img src="https://static.igem.org/mediawiki/parts/7/79/Rumus.JPG" width="300"/><p><br />
<div style="text-align: center;"><br />
<p><img src="https://static.igem.org/mediawiki/parts/f/fb/Graph.JPG" width="500"/></p><br />
<p><small>Figure 1. Ethylene Glycol Concentration LC ITB and LC UC Davis<br />
</small></p><br />
</div><br />
<p>Each sample used for measure the absorbance contain a fixed amount of chromic acid. The absorbance of the chromic acid will represent the total amount of ethylene glycol inversely. It means that if the absorbance is high, the amount of ethylene glycol in solution is low, and if the absorbance is low, the amount of ethylene glycol in solution is high. If the absorbance is high, the amount of chromic acid is high, which means that the amount of ethylene glycol being oxidized by chromic acid is low and vice versa. Both of LC Cutinase UC Davis and OmpA-LC Cutinase ITB_Indonesia shown the degrading activity of PET become ethylene glycol as once of the product.The optimum time to produce ethylene glycol is on the fourth hour after inoculation. </p><br />
<p> Reference</p><br />
<p>Ribitsch D, Enrique HA, Katrin G, Anita D, Sabine Z, Annemarie M, Rosario DR, Georg S, Karl G, Helmut S and Georg MG. 2012. A New Esterase from Thermobifida halotolerans Hydrolyses Polyethylene Terephthalate (PET) and Polylactic Acid (PLA). Polymers. 4: 617-629 </p><br />
<p>Sarker M, Aminul K, Mohammad MR, Mohammed M and ASMD Mohammad. 2011. Waste Polyethylene Terephthalate (PETE-1) Conversion into Liquid Fuel. Journal of Fundamentals of Renewable Energy and Applications. 1</p><br />
<p>Venkatachalam S, Shilpa GN, Jayprakash VL, Prashant RG, Krishna R and Anil KK. 2012. Degradation and Recyclability of Poly (Ethylene Terephthalate). Intech</p><br />
<p><h3> Scaning Electron Microscope (SEM) Result Analysis </h3></p><br />
<p>For biodegrading experiments, we used 3x5 cm2 bottle plastics with average weight 0.05 g. Then, we washed it several times with water and ethanol. </p><br />
<p>Plastic sample was applying just before we inoculated the bacteria on the medium. Bacterial culture were grown at 37C in Luria-Broth medium. After 3 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss. </p><br />
<p>Meanwhile, for UC Davis bacterial culture, we inoculated the bacteria in Luria-Broth medium supplemented with 170 ppm cloramphenicol. After an absorbance A600 nm of 0.6 was reached, we added 1% of arabinose and plastic sample. After 2 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss.</p><br />
<p>We used medium supplemented with antibiotics and plastic sample as control on this experiment. The scaning electron microsccopy analysis of fractured surface of PET film was carried out using Scaning electron microscope. The surface of the treated PET samples were coated with conductive heavy metals such as gold/palladium. </p><br />
<p>SEM image shows that cracks were observed at the surface of a plastic sample (PET) after incubation on the bacterial culture. But, there is no significant weight decreased of the plastics samples. From this SEM study we conclude that both our LC Cut UC Davis and OmpA-LC-Cut Fussion ITB 2014 able to degrade PET plastics samples. Figure 1. shown the control sampel that we could not observe any cracks. Figure 2 and 3 shown the PET degradation by Lc cutinase from UC davis and from ITB_Indonesia. </p><br />
<br />
<br />
<br />
<br />
<br />
<br></div>
Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Data
Team:ITB Indonesia/Data
2014-10-15T15:56:49Z
<p>Erawijantari: </p>
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-modeling">MODELING</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Kids">FUN WITH KIDS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Data</h1><br />
<h3>Ethylene Glycol Assay using Chromic Acid</h3><br />
<p>Poly(ethylene terephthalate) (PET) is a plastic that is a polymer of ethylene terephthalate (C10H8O4) units (Sarker et al., 2010). Polyethylene terephthalate (PET) can be degrade via limited enzymatic hydrolysis of the ester bond of the polymer backbone, one enzyme that have been assesed for this purpose is cutinase (Ribitsch et al., 2012). When polyethylene terephthalate degraded, it will produce terephthalic acid and ethylene glycol (Venkatachalam et al., 2012). Ethylene glycol is one of alcohol compound. Oxidation of alcohols by strong oxidants such as chromic acid (K2Cr2O7) in suphuric acid(H2SO4) is possible, but differs depending on the degree of alcohol. If a reaction has occurred using chromic acid in sulphuric acid, there is a color change from orange to green. In our project we use chromic acid to detect ethylene glycol as product of the PET degradation by LC cutinase enzyme.figure 1 shown the result of ethylene glycol assay using chromic acid.</p><br />
<p><img src="https://static.igem.org/mediawiki/parts/7/79/Rumus.JPG" width="300"/><p><br />
<div style="text-align: center;"><br />
<p><img src="https://static.igem.org/mediawiki/parts/f/fb/Graph.JPG" width="500"/></p><br />
<p><small>Figure 1. Ethylene Glycol Concentration LC ITB and LC UC Davis<br />
</small></p><br />
</div><br />
<p>Each sample used for measure the absorbance contain a fixed amount of chromic acid. The absorbance of the chromic acid will represent the total amount of ethylene glycol inversely. It means that if the absorbance is high, the amount of ethylene glycol in solution is low, and if the absorbance is low, the amount of ethylene glycol in solution is high. If the absorbance is high, the amount of chromic acid is high, which means that the amount of ethylene glycol being oxidized by chromic acid is low and vice versa. Both of LC Cutinase UC Davis and OmpA-LC Cutinase ITB_Indonesia shown the degrading activity of PET become ethylene glycol as once of the product.The optimum time to produce ethylene glycol is on the fourth hour after inoculation. </p><br />
<p> Reference</p><br />
<p>Ribitsch D, Enrique HA, Katrin G, Anita D, Sabine Z, Annemarie M, Rosario DR, Georg S, Karl G, Helmut S and Georg MG. 2012. A New Esterase from Thermobifida halotolerans Hydrolyses Polyethylene Terephthalate (PET) and Polylactic Acid (PLA). Polymers. 4: 617-629 </p><br />
<p>Sarker M, Aminul K, Mohammad MR, Mohammed M and ASMD Mohammad. 2011. Waste Polyethylene Terephthalate (PETE-1) Conversion into Liquid Fuel. Journal of Fundamentals of Renewable Energy and Applications. 1</p><br />
<p>Venkatachalam S, Shilpa GN, Jayprakash VL, Prashant RG, Krishna R and Anil KK. 2012. Degradation and Recyclability of Poly (Ethylene Terephthalate). Intech</p><br />
<p><h3> Scaning Electron Microscope (SEM) Result Analysis </h3></p><br />
<p>For biodegrading experiments, we used 3x5 cm2 bottle plastics with average weight 0.05 g. Then, we washed it several times with water and ethanol. </p><br />
<p>Plastic sample was applying just before we inoculated the bacteria on the medium. Bacterial culture were grown at 37C in Luria-Broth medium. After 3 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss. </p><br />
<p>Meanwhile, for UC Davis bacterial culture, we inoculated the bacteria in Luria-Broth medium supplemented with 170 ppm cloramphenicol. After an absorbance A600 nm of 0.6 was reached, we added 1% of arabinose and plastic sample. After 2 days of incubation, we washed the plastic with water and ethanol then dried for measurement of the weight loss.</p><br />
<p>We used medium supplemented with antibiotics and plastic sample as control on this experiment. The scaning electron microsccopy analysis of fractured surface of PET film was carried out using Scaning electron microscope. The surface of the treated PET samples were coated with conductive heavy metals such as gold/palladium. </p><br />
<p>SEM image shows that cracks were observed at the surface of a plastic sample (PET) after incubation on the bacterial culture. But, there is no significant weight decreased of the plastics samples. From this SEM study we conclude that both our LC Cut UC Davis and OmpA-LC-Cut Fussion ITB 2014 able to degrade PET plastics samples. Figure 1. shown the control sampel that we could not observe any cracks. Figure 2 and 3 shown the PET degradation by Lc cutinase from UC davis and from ITB_Indonesia. </p><br />
<br />
<br />
<br />
<br />
<br />
<br></div>
Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Data
Team:ITB Indonesia/Data
2014-10-15T15:53:43Z
<p>Erawijantari: </p>
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</ul><br />
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<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Data</h1><br />
<h3>Ethylene Glycol Assay using Chromic Acid</h3><br />
<p>Poly(ethylene terephthalate) (PET) is a plastic that is a polymer of ethylene terephthalate (C10H8O4) units (Sarker et al., 2010). Polyethylene terephthalate (PET) can be degrade via limited enzymatic hydrolysis of the ester bond of the polymer backbone, one enzyme that have been assesed for this purpose is cutinase (Ribitsch et al., 2012). When polyethylene terephthalate degraded, it will produce terephthalic acid and ethylene glycol (Venkatachalam et al., 2012). Ethylene glycol is one of alcohol compound. Oxidation of alcohols by strong oxidants such as chromic acid (K2Cr2O7) in suphuric acid(H2SO4) is possible, but differs depending on the degree of alcohol. If a reaction has occurred using chromic acid in sulphuric acid, there is a color change from orange to green. In our project we use chromic acid to detect ethylene glycol as product of the PET degradation by LC cutinase enzyme.figure 1 shown the result of ethylene glycol assay using chromic acid.</p><br />
<p><img src="https://static.igem.org/mediawiki/parts/7/79/Rumus.JPG" width="300"/><p><br />
<div style="text-align: center;"><br />
<p><img src="https://static.igem.org/mediawiki/parts/f/fb/Graph.JPG" width="500"/></p><br />
<p><small>Figure 1. Ethylene Glycol Concentration LC ITB and LC UC Davis<br />
</small></p><br />
</div><br />
<p>Each sample used for measure the absorbance contain a fixed amount of chromic acid. The absorbance of the chromic acid will represent the total amount of ethylene glycol inversely. It means that if the absorbance is high, the amount of ethylene glycol in solution is low, and if the absorbance is low, the amount of ethylene glycol in solution is high. If the absorbance is high, the amount of chromic acid is high, which means that the amount of ethylene glycol being oxidized by chromic acid is low and vice versa. Both of LC Cutinase UC Davis and OmpA-LC Cutinase ITB_Indonesia shown the degrading activity of PET become ethylene glycol as once of the product.The optimum time to produce ethylene glycol is on the fourth hour after inoculation. </p><br />
<p> Reference</p><br />
<p>Ribitsch D, Enrique HA, Katrin G, Anita D, Sabine Z, Annemarie M, Rosario DR, Georg S, Karl G, Helmut S and Georg MG. 2012. A New Esterase from Thermobifida halotolerans Hydrolyses Polyethylene Terephthalate (PET) and Polylactic Acid (PLA). Polymers. 4: 617-629 </p><br />
<p>Sarker M, Aminul K, Mohammad MR, Mohammed M and ASMD Mohammad. 2011. Waste Polyethylene Terephthalate (PETE-1) Conversion into Liquid Fuel. Journal of Fundamentals of Renewable Energy and Applications. 1</p><br />
<p>Venkatachalam S, Shilpa GN, Jayprakash VL, Prashant RG, Krishna R and Anil KK. 2012. Degradation and Recyclability of Poly (Ethylene Terephthalate). Intech</p><br />
<p><h3> Scaning Electron Microscope (SEM) Result Analysis </h3></p><br />
<br />
<br />
<br />
<br />
<br />
<br></div>
Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Data
Team:ITB Indonesia/Data
2014-10-15T15:51:39Z
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SelfMod">SELF REGULATORY MODULE</a></li><br />
</ul><br />
</li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Modeling">MODELING</a></li><br />
<li>WETLAB<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/protocol">PROTOCOL</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Data">DATA</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/ProteinModel">PROTEIN MODEL</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Achievement">ACHIEVEMENT</a></li><br />
</ul><br />
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<li>NOTEBOOK<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-modeling">MODELING</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/nb-wetlab">WETLAB</a></li><br />
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<li>HUMAN PRACTICE<br />
<ul><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/Unpas">INTRO SYNBIO TO UNPAS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynBio">SYNBIO CLASS</a></li><br />
<li><a href="https://2014.igem.org/Team:ITB_Indonesia/SynbiGreen">SYNBIGREEN SURVEY</a></li><br />
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<li><a href="https://2014.igem.org/Team:ITB_Indonesia/UPI">SHARING SYNBIO IN UPI</a></li><br />
</ul><br />
</li><br />
</ul><br />
</div><br />
<br />
<br><br />
<h1>Data</h1><br />
<h3>Ethylene Glycol Assay using Chromic Acid</h3><br />
<p>Poly(ethylene terephthalate) (PET) is a plastic that is a polymer of ethylene terephthalate (C10H8O4) units (Sarker et al., 2010). Polyethylene terephthalate (PET) can be degrade via limited enzymatic hydrolysis of the ester bond of the polymer backbone, one enzyme that have been assesed for this purpose is cutinase (Ribitsch et al., 2012). When polyethylene terephthalate degraded, it will produce terephthalic acid and ethylene glycol (Venkatachalam et al., 2012). Ethylene glycol is one of alcohol compound. Oxidation of alcohols by strong oxidants such as chromic acid (K2Cr2O7) in suphuric acid(H2SO4) is possible, but differs depending on the degree of alcohol. If a reaction has occurred using chromic acid in sulphuric acid, there is a color change from orange to green. In our project we use chromic acid to detect ethylene glycol as product of the PET degradation by LC cutinase enzyme.figure 1 shown the result of ethylene glycol assay using chromic acid.</p><br />
<p><img src="https://static.igem.org/mediawiki/parts/7/79/Rumus.JPG" width="300"/><p><br />
<div style="text-align: center;"><br />
<p><img src="https://static.igem.org/mediawiki/parts/f/fb/Graph.JPG" width="500"/></p><br />
<p><small>Figure 1. Ethylene Glycol Concentration LC ITB and LC UC Davis<br />
</small></p><br />
</div><br />
<p>Each sample used for measure the absorbance contain a fixed amount of chromic acid. The absorbance of the chromic acid will represent the total amount of ethylene glycol inversely. It means that if the absorbance is high, the amount of ethylene glycol in solution is low, and if the absorbance is low, the amount of ethylene glycol in solution is high. If the absorbance is high, the amount of chromic acid is high, which means that the amount of ethylene glycol being oxidized by chromic acid is low and vice versa. Both of LC Cutinase UC Davis and OmpA-LC Cutinase ITB_Indonesia shown the degrading activity of PET become ethylene glycol as once of the product.The optimum time to produce ethylene glycol is on the fourth hour after inoculation. </p><br />
<p> Reference</p><br />
<p>Ribitsch D, Enrique HA, Katrin G, Anita D, Sabine Z, Annemarie M, Rosario DR, Georg S, Karl G, Helmut S and Georg MG. 2012. A New Esterase from Thermobifida halotolerans Hydrolyses Polyethylene Terephthalate (PET) and Polylactic Acid (PLA). Polymers. 4: 617-629 </p><br />
<p>Sarker M, Aminul K, Mohammad MR, Mohammed M and ASMD Mohammad. 2011. Waste Polyethylene Terephthalate (PETE-1) Conversion into Liquid Fuel. Journal of Fundamentals of Renewable Energy and Applications. 1</p><br />
<p>Venkatachalam S, Shilpa GN, Jayprakash VL, Prashant RG, Krishna R and Anil KK. 2012. Degradation and Recyclability of Poly (Ethylene Terephthalate). Intech</p><br />
<br />
<br />
<br />
<br />
<br />
<br></div>
Erawijantari
http://2014.igem.org/Team:ITB_Indonesia/Data
Team:ITB Indonesia/Data
2014-10-15T15:49:56Z
<p>Erawijantari: </p>
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<br><br />
<h1>Data</h1><br />
<h3>Ethylene Glycol Assay using Chromic Acid</h3><br />
<p>Poly(ethylene terephthalate) (PET) is a plastic that is a polymer of ethylene terephthalate (C10H8O4) units (Sarker et al., 2010). Polyethylene terephthalate (PET) can be degrade via limited enzymatic hydrolysis of the ester bond of the polymer backbone, one enzyme that have been assesed for this purpose is cutinase (Ribitsch et al., 2012). When polyethylene terephthalate degraded, it will produce terephthalic acid and ethylene glycol (Venkatachalam et al., 2012). Ethylene glycol is one of alcohol compound. Oxidation of alcohols by strong oxidants such as chromic acid (K2Cr2O7) in suphuric acid(H2SO4) is possible, but differs depending on the degree of alcohol. If a reaction has occurred using chromic acid in sulphuric acid, there is a color change from orange to green. In our project we use chromic acid to detect ethylene glycol as product of the PET degradation by LC cutinase enzyme.figure 1 shown the result of ethylene glycol assay using chromic acid.</p><br />
<p><img src="https://static.igem.org/mediawiki/parts/7/79/Rumus.JPG" width="300"/><p><br />
<p><img src="https://static.igem.org/mediawiki/parts/f/fb/Graph.JPG" width="500"/></p><br />
<p><small>Figure 1. Ethylene Glycol Concentration LC ITB and LC UC Davis<br />
</small></p><br />
<p>Each sample used for measure the absorbance contain a fixed amount of chromic acid. The absorbance of the chromic acid will represent the total amount of ethylene glycol inversely. It means that if the absorbance is high, the amount of ethylene glycol in solution is low, and if the absorbance is low, the amount of ethylene glycol in solution is high. If the absorbance is high, the amount of chromic acid is high, which means that the amount of ethylene glycol being oxidized by chromic acid is low and vice versa. Both of LC Cutinase UC Davis and OmpA-LC Cutinase ITB_Indonesia shown the degrading activity of PET become ethylene glycol as once of the product.The optimum time to produce ethylene glycol is on the fourth hour after inoculation. </p><br />
<p> Reference</p><br />
<p>Ribitsch D, Enrique HA, Katrin G, Anita D, Sabine Z, Annemarie M, Rosario DR, Georg S, Karl G, Helmut S and Georg MG. 2012. A New Esterase from Thermobifida halotolerans Hydrolyses Polyethylene Terephthalate (PET) and Polylactic Acid (PLA). Polymers. 4: 617-629 </p><br />
<p>Sarker M, Aminul K, Mohammad MR, Mohammed M and ASMD Mohammad. 2011. Waste Polyethylene Terephthalate (PETE-1) Conversion into Liquid Fuel. Journal of Fundamentals of Renewable Energy and Applications. 1</p><br />
<p>Venkatachalam S, Shilpa GN, Jayprakash VL, Prashant RG, Krishna R and Anil KK. 2012. Degradation and Recyclability of Poly (Ethylene Terephthalate). Intech</p><br />
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