"
Team:UI-Indonesia/Parts/Characterization
From 2014.igem.org
| Line 47: | Line 47: | ||
To examine in which concentration of IPTG that can be toxic for E.coli as bacterial vector | To examine in which concentration of IPTG that can be toxic for E.coli as bacterial vector | ||
<br><br> | <br><br> | ||
| + | <b>Result</b><br><br> | ||
| + | <img src="https://static.igem.org/mediawiki/2014/9/90/Antimicrobial_result.png" width="auto" height="auto"> | ||
| + | <img src="https://static.igem.org/mediawiki/2014/6/64/Antimicrobial_assay.png" width="auto" height="auto"> | ||
| + | <br><br> | ||
| + | |||
| + | <b>Discussion</b><br> | ||
| + | |||
| + | |||
| + | </p> | ||
| + | </div> | ||
| + | |||
| + | |||
| + | <div id="page-box1"> | ||
| + | <h2>Alpha amylase and Nuclease Characterization in Biofilm Removal</h2> | ||
| + | <p> | ||
| + | <b>Background</b><br> | ||
| + | Biofilm as matrix extracelullar polymeric substances (EPS) causes an increase in antibiotic resistance and pathogenecity of pathogenic bacteria. | ||
| + | <br><br> | ||
| + | |||
| + | <b>Aim</b><br> | ||
| + | We present our Genius E.coli agent that can degrade EPS biofilm by secreting enzymes, such as alpha amylase and nuclease. | ||
| + | <br><br> | ||
| + | |||
| + | <b>Methods</b><br> | ||
| + | We designed an E. coli having inserted MalS and HlyA gene in pSB1C3 plasmid in E.coli Top10 Bba_K13440004 (MalS HlyA in pSB1C3), and E. coli having plasmid inserted MalS and HlyA gene in pSB1C3 under control strong prommoter constitutive 100 Bba_K13440005 (Sp100 MalS HlyA in pSB1C3) and E. coli top10 wild type as negative control were growth on LB starch agar. <br> | ||
| + | Mals is gene that has responsible to encode the enzyme expression of alpha amylase for E. coli strain K12, the length is 1967 bp. The HlyA is a signal peptide found in the C-terminal signal sequence of alpha-hemolysin (HlyA). Fusion of the HlyA signal peptide to the target protein (MalS) cause the excretion of protein to extracellular medium in a single step. | ||
| + | <br> | ||
| + | Upon successful cloning of the three genes into our E.coli, we continued to confirm that all three genes are required have hydrolizing amilum activity on LB starch agar. We using iodine test to staining the amilum, and see the clearzone for some time variables. | ||
| + | <br><br> | ||
| + | |||
<b>Result</b><br><br> | <b>Result</b><br><br> | ||
<img src="https://static.igem.org/mediawiki/2014/9/90/Antimicrobial_result.png" width="auto" height="auto"> | <img src="https://static.igem.org/mediawiki/2014/9/90/Antimicrobial_result.png" width="auto" height="auto"> | ||
Revision as of 23:05, 17 October 2014
Peptide 1018 Antimicrobial Assay
Background
Peptide 1018 is small cationic peptide under T5 promoter (inducible promoter). Peptide 1018 in particular concentration can be toxic for bacterial vector. Isopropyl thiogalactoside (IPTG) is analogue of lactose that is able to induce activation of T5 promoter to start transcription.
Aim
To examine in which concentration of IPTG that can be toxic for E.coli as bacterial vector
Result
Discussion
Alpha amylase and Nuclease Characterization in Biofilm Removal
Background
Biofilm as matrix extracelullar polymeric substances (EPS) causes an increase in antibiotic resistance and pathogenecity of pathogenic bacteria.
Aim
We present our Genius E.coli agent that can degrade EPS biofilm by secreting enzymes, such as alpha amylase and nuclease.
Methods
We designed an E. coli having inserted MalS and HlyA gene in pSB1C3 plasmid in E.coli Top10 Bba_K13440004 (MalS HlyA in pSB1C3), and E. coli having plasmid inserted MalS and HlyA gene in pSB1C3 under control strong prommoter constitutive 100 Bba_K13440005 (Sp100 MalS HlyA in pSB1C3) and E. coli top10 wild type as negative control were growth on LB starch agar.
Mals is gene that has responsible to encode the enzyme expression of alpha amylase for E. coli strain K12, the length is 1967 bp. The HlyA is a signal peptide found in the C-terminal signal sequence of alpha-hemolysin (HlyA). Fusion of the HlyA signal peptide to the target protein (MalS) cause the excretion of protein to extracellular medium in a single step.
Upon successful cloning of the three genes into our E.coli, we continued to confirm that all three genes are required have hydrolizing amilum activity on LB starch agar. We using iodine test to staining the amilum, and see the clearzone for some time variables.
Result
Discussion
