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Team:Toulouse/Result/parts/Submitted parts
From 2014.igem.org
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text-align:justify; | text-align:justify; | ||
vertical-align:top; | vertical-align:top; | ||
| - | border-width: | + | border-width:0px; |
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<table> | <table> | ||
<td valign="top"; align="center";> | <td valign="top"; align="center";> | ||
| - | <p class="title2"><a href="http://parts.igem.org/Part:BBa_K1364000">BBa_K1364000</a></p></td> | + | <p class="title2"; "line-height:6px"; "padding:10px";> |
| + | <a href="http://parts.igem.org/Part:BBa_K1364000">BBa_K1364000</a></p></td> | ||
<td valign="top"; align="center";> | <td valign="top"; align="center";> | ||
<div class="technology2">N-acetylglucosamine based chemotaxis for <i>Bacillus subtilis</i></div> | <div class="technology2">N-acetylglucosamine based chemotaxis for <i>Bacillus subtilis</i></div> | ||
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these domains correspond to the amino acids 1-37 and 282-610. | these domains correspond to the amino acids 1-37 and 282-610. | ||
The extracellular domain of VCD is inserted between the intracellular regions of McpA | The extracellular domain of VCD is inserted between the intracellular regions of McpA | ||
| - | to sense N-acetylglucosamine and corresponds to the amino acids 31-310. | + | to sense N-acetylglucosamine and corresponds to the amino acids 31-310.</p> |
<img src="https://static.igem.org/mediawiki/2014/a/a2/BBa_K1364000.png"> | <img src="https://static.igem.org/mediawiki/2014/a/a2/BBa_K1364000.png"> | ||
<p class="title4">Type</p> | <p class="title4">Type</p> | ||
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<p class="texte">Uniprot:<br> | <p class="texte">Uniprot:<br> | ||
- <a href="http://www.uniprot.org/uniprot/P39214">McpA</a><br> | - <a href="http://www.uniprot.org/uniprot/P39214">McpA</a><br> | ||
| - | - <a href="http://www.uniprot.org/uniprot/C3NYT2">N-acetylglucosamine regulated methyl-accepting chemotaxis protein</a | + | - <a href="http://www.uniprot.org/uniprot/C3NYT2">N-acetylglucosamine regulated methyl-accepting chemotaxis protein</a></p> |
<!--à compléter--> | <!--à compléter--> | ||
| - | |||
| - | |||
<p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 0); return false">Collapse</a></p> | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 0); return false">Collapse</a></p> | ||
</div></td></table> | </div></td></table> | ||
| - | < | + | <table> |
| - | <div class=" | + | <td valign="top"; align="center";> |
| - | + | <p class="title2"; "line-height:6px"; "padding:10px";> | |
| - | + | <a href="http://parts.igem.org/Part:BBa_K1364004">BBa_K1364004</a></p></td> | |
| + | <td valign="top"; align="center";> | ||
| + | <div class="technology2">P<sub>veg</sub> + N-acetylglucosamine based chemotaxis for <i>Bacillus subtilis</i></div> | ||
| + | <div class="thelanguage2"> | ||
<p class="texte">This part is designed to enable a N-acetylglucosamine based chemotaxis in <i>Bacillus subtilis</i>.</p> | <p class="texte">This part is designed to enable a N-acetylglucosamine based chemotaxis in <i>Bacillus subtilis</i>.</p> | ||
<p class="title4">Design</p> | <p class="title4">Design</p> | ||
<p class="texte">This expression cassette is designed for the expression of an antifungal peptide, | <p class="texte">This expression cassette is designed for the expression of an antifungal peptide, | ||
| - | D4E1 and for its secretion in <i> | + | D4E1 and for its secretion in <i>B. subtilis</i>. |
It is composed of the strong, constitutive promoter of <i>B. subtilis</i> | It is composed of the strong, constitutive promoter of <i>B. subtilis</i> | ||
P<sub>veg</sub> (<a href="http://parts.igem.org/wiki/index.php/Part:BBa_K823003">K823003</a>), | P<sub>veg</sub> (<a href="http://parts.igem.org/wiki/index.php/Part:BBa_K823003">K823003</a>), | ||
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- <a href="http://www.uniprot.org/uniprot/C3NYT2">N-acetylglucosamine regulated methyl-accepting chemotaxis protein</a></p> | - <a href="http://www.uniprot.org/uniprot/C3NYT2">N-acetylglucosamine regulated methyl-accepting chemotaxis protein</a></p> | ||
| - | + | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 1); return false">Collapse</a></p> | |
| - | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone(' | + | </div></td></table> |
| - | </div> | + | |
<br></br> | <br></br> | ||
| - | <p class=" | + | <p class="title1">Binding</p> |
| - | + | <table> | |
| - | < | + | <td valign="top"; align="center";> |
| - | <div class=" | + | <p class="title2"; "line-height:6px"; "padding:10px";> |
| + | <a href="http://parts.igem.org/Part:BBa_K1364005">BBa_K1364005</a></p></td> | ||
| + | <td valign="top"; align="center";> | ||
| + | <div class="technology2"></a>P<sub>veg</sub> + Chitin Binding Cell Wall protein</div> | ||
| + | <div class="thelanguage2"> | ||
<p class="texte">This part is designed to enable the binding of <i>Bacillus subtilis</i> to the fungi wall made of chitin.<p/> | <p class="texte">This part is designed to enable the binding of <i>Bacillus subtilis</i> to the fungi wall made of chitin.<p/> | ||
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<p class="title4">References</p> | <p class="title4">References</p> | ||
<p class="texte">- Wong E, Vaaje-Kolstad G, Ghosh A, Hurtado-Guerrero R, Konarev PV, et al. (2012)<br> | <p class="texte">- Wong E, Vaaje-Kolstad G, Ghosh A, Hurtado-Guerrero R, Konarev PV, et al. (2012)<br> | ||
| - | - The Vibrio cholerae Colonization Factor GbpA Possesses a Modular Structure that Governs Binding to Different Host Surfaces. PLoS Pathog 8(1): e1002373. doi:10.1371/journal.ppat.1002373 | + | - The Vibrio cholerae Colonization Factor GbpA Possesses a Modular Structure that Governs Binding to Different Host Surfaces. PLoS Pathog 8(1): e1002373. doi:10.1371/journal.ppat.1002373 |
</p> | </p> | ||
| - | + | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 2); return false">Collapse</a></p> | |
| - | + | </div></td></table> | |
| - | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone(' | + | |
| - | </div> | + | |
<br></br> | <br></br> | ||
| - | <p class=" | + | <p class="title1">Fungicides</p> |
<p class="title3">GAFP-1</p> | <p class="title3">GAFP-1</p> | ||
<br> | <br> | ||
| - | + | <table> | |
| - | < | + | <td valign="top"; align="center";> |
| - | <div class=" | + | <p class="title2"; "line-height:6px"; "padding:10px";> |
| + | <a href="http://parts.igem.org/Part:BBa_K1364002">BBa_K1364002</a></p></td> | ||
| + | <td valign="top"; align="center";> | ||
| + | <div class="technology2"> RBS - Antifungal GAFP-1</div> | ||
| + | <div class="thelanguage2"> | ||
<p class="texte">The <i>Gastrodia</i> anti-fungal protein(GAFP-1), also known as gastrodianin, is a mannose and chitin binding lectin | <p class="texte">The <i>Gastrodia</i> anti-fungal protein(GAFP-1), also known as gastrodianin, is a mannose and chitin binding lectin | ||
| - | originating from the Asiatic orchid Gastrodia elata, a traditional Chinese medicinal herb cultured for thousands of years. | + | originating from the Asiatic orchid <i>Gastrodia elata</i>, a traditional Chinese medicinal herb cultured for thousands of years. |
GAFP-1 is composed of 15 amino acids LDSLSFSYNNFEEDD and is able to inhibit the growth of multiples species of plant pathogenic fungi.</p> | GAFP-1 is composed of 15 amino acids LDSLSFSYNNFEEDD and is able to inhibit the growth of multiples species of plant pathogenic fungi.</p> | ||
<p class="title4">Design</p> | <p class="title4">Design</p> | ||
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<p class="title4">References</p> | <p class="title4">References</p> | ||
<p class="texte">- Wong E, Vaaje-Kolstad G, Ghosh A, Hurtado-Guerrero R, Konarev PV, et al. (2012)<br> | <p class="texte">- Wong E, Vaaje-Kolstad G, Ghosh A, Hurtado-Guerrero R, Konarev PV, et al. (2012)<br> | ||
| - | - The Vibrio cholerae Colonization Factor GbpA Possesses a Modular Structure that Governs Binding to Different Host Surfaces. PLoS Pathog 8(1): e1002373. doi:10.1371/journal.ppat.1002373 | + | - The Vibrio cholerae Colonization Factor GbpA Possesses a Modular Structure that Governs Binding to Different Host Surfaces. PLoS Pathog 8(1): e1002373. doi:10.1371/journal.ppat.1002373</p> |
| - | + | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 3); return false">Collapse</a></p> | |
| - | + | </div></td></table> | |
| - | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone(' | + | |
| - | </div> | + | |
| - | + | ||
| - | < | + | |
| - | + | ||
| - | + | ||
| + | <table> | ||
| + | <td valign="top"; align="center";> | ||
| + | <p class="title2"; "line-height:6px"; "padding:10px";> | ||
| + | <a href="http://parts.igem.org/Part:BBa_K1364007">BBa_K1364007</a></p></td> | ||
| + | <td valign="top"; align="center";> | ||
| + | <div class="technology2">RBS + Antifungal GAFP-1 + Double terminator</div> | ||
| + | <div class="thelanguage2"> | ||
<p class="texte">The <i>Gastrodia</i> anti-fungal protein(GAFP-1), also known as gastrodianin, is a mannose and chitin binding lectin | <p class="texte">The <i>Gastrodia</i> anti-fungal protein(GAFP-1), also known as gastrodianin, is a mannose and chitin binding lectin | ||
| - | originating from the Asiatic orchid Gastrodia elata, a traditional Chinese medicinal herb cultured for thousands of years. | + | originating from the Asiatic orchid <i>Gastrodia elata</i>, a traditional Chinese medicinal herb cultured for thousands of years. |
GAFP-1 is composed of 15 amino acids LDSLSFSYNNFEEDD and is able to inhibit the growth of multiples species of plant pathogenic fungi.</p> | GAFP-1 is composed of 15 amino acids LDSLSFSYNNFEEDD and is able to inhibit the growth of multiples species of plant pathogenic fungi.</p> | ||
<p class="title4">Design</p> | <p class="title4">Design</p> | ||
| Line 297: | Line 305: | ||
<p class="title4">References</p> | <p class="title4">References</p> | ||
<p class="texte">- Wong E, Vaaje-Kolstad G, Ghosh A, Hurtado-Guerrero R, Konarev PV, et al. (2012)<br> | <p class="texte">- Wong E, Vaaje-Kolstad G, Ghosh A, Hurtado-Guerrero R, Konarev PV, et al. (2012)<br> | ||
| - | - The Vibrio cholerae Colonization Factor GbpA Possesses a Modular Structure that Governs Binding to Different Host Surfaces. PLoS Pathog 8(1): e1002373. doi:10.1371/journal.ppat.1002373 | + | - The Vibrio cholerae Colonization Factor GbpA Possesses a Modular Structure that Governs Binding to Different Host Surfaces. PLoS Pathog 8(1): e1002373. doi:10.1371/journal.ppat.1002373</p> |
| - | + | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 4); return false">Collapse</a></p> | |
| - | + | </div></td></table> | |
| - | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone(' | + | |
| - | </div> | + | |
| - | + | ||
| - | < | + | |
| - | + | ||
| + | <table> | ||
| + | <td valign="top";align="center";> | ||
| + | <p class="title2"; "line-height:6px"; "padding:10px";> | ||
| + | <a href="http://parts.igem.org/Part:BBa_K1364008">BBa_K1364008</a></p></td> | ||
| + | <td valign="top"; align="center";> | ||
| + | <div class="technology2"> P<sub>veg</sub> - strong RBS - Antifungal GAFP-1 - Double terminator</div> | ||
| + | <div class="thelanguage2"> | ||
<p class="texte">This expression cassette is designed for the expression and secretion of the <i>Gastrodia</i> anti-fungal protein(GAFP-1), also known as gastrodianin. GAFP-1 is a mannose and chitin binding lectin | <p class="texte">This expression cassette is designed for the expression and secretion of the <i>Gastrodia</i> anti-fungal protein(GAFP-1), also known as gastrodianin. GAFP-1 is a mannose and chitin binding lectin | ||
| - | originating from the Asiatic orchid Gastrodia elata, a traditional Chinese medicinal herb cultured for thousands of years. | + | originating from the Asiatic orchid <i>Gastrodia elata</i>, a traditional Chinese medicinal herb cultured for thousands of years. |
GAFP-1 is composed of 15 amino acids LDSLSFSYNNFEEDD and is able to inhibit the growth of multiples species of plant pathogenic fungi.</p> | GAFP-1 is composed of 15 amino acids LDSLSFSYNNFEEDD and is able to inhibit the growth of multiples species of plant pathogenic fungi.</p> | ||
<p class="title4">Design</p> | <p class="title4">Design</p> | ||
| Line 314: | Line 324: | ||
the open reading frame of the <i>Gastrodia</i> anti-fungal protein 1 (GAFP-1) | the open reading frame of the <i>Gastrodia</i> anti-fungal protein 1 (GAFP-1) | ||
and a double terminator (<a href="http://parts.igem.org/Part:BBa_B0015">B0015</a>). | and a double terminator (<a href="http://parts.igem.org/Part:BBa_B0015">B0015</a>). | ||
| - | optimized for its expression and its secretion in <i> | + | optimized for its expression and its secretion in <i>B. subtilis</i>. |
The codon optimization was made thanks to the DNA 2.0 software program.</p> | The codon optimization was made thanks to the DNA 2.0 software program.</p> | ||
<img src="https://static.igem.org/mediawiki/2014/6/67/BBa_K1364008.png"> | <img src="https://static.igem.org/mediawiki/2014/6/67/BBa_K1364008.png"> | ||
| Line 323: | Line 333: | ||
<p class="title4">References</p> | <p class="title4">References</p> | ||
<p class="texte">- Wong E, Vaaje-Kolstad G, Ghosh A, Hurtado-Guerrero R, Konarev PV, et al. (2012)<br> | <p class="texte">- Wong E, Vaaje-Kolstad G, Ghosh A, Hurtado-Guerrero R, Konarev PV, et al. (2012)<br> | ||
| - | - The Vibrio cholerae Colonization Factor GbpA Possesses a Modular Structure that Governs Binding to Different Host Surfaces. PLoS Pathog 8(1): e1002373. doi:10.1371/journal.ppat.1002373 | + | - The Vibrio cholerae Colonization Factor GbpA Possesses a Modular Structure that Governs Binding to Different Host Surfaces. PLoS Pathog 8(1): e1002373. doi:10.1371/journal.ppat.1002373 |
</p> | </p> | ||
| - | + | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 5); return false">Collapse</a></p> | |
| - | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone(' | + | </div></td></table> |
| - | </div> | + | |
<br></br> | <br></br> | ||
| Line 334: | Line 343: | ||
<br> | <br> | ||
| - | + | <table> | |
| - | < | + | <td valign="top"; align="center";> |
| - | <div class=" | + | <p class="title2"; "line-height:6px"; "padding:10px";> |
| - | + | <a href="http://parts.igem.org/Part:BBa_K1364003">BBa_K1364003</a></p></td> | |
| - | + | <td valign="top"; align="center";> | |
| + | <div class="technology2">RBS - Antifungal D4E1 - Double terminator</div> | ||
| + | <div class="thelanguage2"> | ||
<p class="texte">D4E1 is a linear synthetic peptide of 17 amino acids | <p class="texte">D4E1 is a linear synthetic peptide of 17 amino acids | ||
which has shown to have antifungal activities | which has shown to have antifungal activities | ||
| Line 347: | Line 358: | ||
a Double terminator (<a href="http://parts.igem.org/Part:BBa_B0015">B0015</a>) | a Double terminator (<a href="http://parts.igem.org/Part:BBa_B0015">B0015</a>) | ||
optimized for its expression and its secretion in <i>Bacillus subtilis</i>. | optimized for its expression and its secretion in <i>Bacillus subtilis</i>. | ||
| - | This part was optimized for the expression and its secretion in <i> | + | This part was optimized for the expression and its secretion in <i>B. subtilis </i> |
thanks to the DNA 2.0 software program.</p> | thanks to the DNA 2.0 software program.</p> | ||
<img src="https://static.igem.org/mediawiki/2014/b/bb/BBa_K1364003.png"> | <img src="https://static.igem.org/mediawiki/2014/b/bb/BBa_K1364003.png"> | ||
| Line 357: | Line 368: | ||
<p class="texte">- De Lucca AJ, Bland JM, Grimm C, Jacks TJ, Cary JW, Jaynes JM, Cleveland TE, Walsh TJ. Fungicidal properties, sterol binding, and proteolytic resistance of the synthetic peptide D4E1.<br> | <p class="texte">- De Lucca AJ, Bland JM, Grimm C, Jacks TJ, Cary JW, Jaynes JM, Cleveland TE, Walsh TJ. Fungicidal properties, sterol binding, and proteolytic resistance of the synthetic peptide D4E1.<br> | ||
- Can J Microbiol. 1998 Jun;44(6):514-20.</p> | - Can J Microbiol. 1998 Jun;44(6):514-20.</p> | ||
| + | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 6); return false">Collapse</a></p> | ||
| + | </div></td></table> | ||
| - | < | + | <table> |
| - | < | + | <td valign="top"; align="center";> |
| - | + | <p class="title2"; "line-height:6px"; "padding:10px";> | |
| - | + | <a href="http://parts.igem.org/Part:BBa_K1364009">BBa_K1364009</a></p></td> | |
| - | + | <td valign="top"; align="center";> | |
| - | <div class=" | + | <div class="technology2">P<sub>veg</sub> - RBS - Antifungal D4E1 - Double Terminator</div> |
| - | + | <div class="thelanguage2"> | |
<p class="texte"> | <p class="texte"> | ||
D4E1 is a linear synthetic peptide of 17 amino acids | D4E1 is a linear synthetic peptide of 17 amino acids | ||
| Line 370: | Line 383: | ||
by complexing with a sterol present in conodial wall of a varety of fungi.</p> | by complexing with a sterol present in conodial wall of a varety of fungi.</p> | ||
<p class="title4">Design</p> | <p class="title4">Design</p> | ||
| - | <p class="texte">This part is composed of the strong, constitutive promoter of <i> | + | <p class="texte">This part is composed of the strong, constitutive promoter of <i>Bacillus subtilis</i> |
P<sub>veg</sub> (<a href="http://parts.igem.org/wiki/index.php/Part:BBa_K823003">K823003</a>), | P<sub>veg</sub> (<a href="http://parts.igem.org/wiki/index.php/Part:BBa_K823003">K823003</a>), | ||
a strong RBS (<a href="http://parts.igem.org/Part:BBa_K780002">K780002</a>), | a strong RBS (<a href="http://parts.igem.org/Part:BBa_K780002">K780002</a>), | ||
the open reading frame of D4E1 and | the open reading frame of D4E1 and | ||
a double terminator (<a href="http://parts.igem.org/Part:BBa_B0015">B0015</a>). | a double terminator (<a href="http://parts.igem.org/Part:BBa_B0015">B0015</a>). | ||
| - | This part was optimized for the expression and its secretion in <i> | + | This part was optimized for the expression and its secretion in <i>B.subtilis </i> |
thanks to the DNA 2.0 software program.</p> | thanks to the DNA 2.0 software program.</p> | ||
<img src="https://static.igem.org/mediawiki/2014/4/47/BBa_K1364009.png"> | <img src="https://static.igem.org/mediawiki/2014/4/47/BBa_K1364009.png"> | ||
| Line 386: | Line 399: | ||
- Can J Microbiol. 1998 Jun;44(6):514-20.</p> | - Can J Microbiol. 1998 Jun;44(6):514-20.</p> | ||
| - | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone(' | + | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 7); return false">Collapse</a></p> |
| - | </div> | + | </div></td></table> |
<br></br> | <br></br> | ||
| Line 393: | Line 406: | ||
<p class="title3">EcAMP-1</p> | <p class="title3">EcAMP-1</p> | ||
| - | < | + | <table> |
| - | <div class=" | + | <td valign="top"; align="center";> |
| - | + | <p class="title2"; "line-height:6px"; "padding:10px";> | |
| + | <a href="http://parts.igem.org/Part:BBa_K1364010">BBa_K1364010</a></p></td> | ||
| + | <td valign="top"; align="center";> | ||
| + | <div class="technology2">P<sub>veg</sub> - SpoVG - Antifungal EcAMP-1</div> | ||
| + | <div class="thelanguage2"> | ||
<p class="texte"> | <p class="texte"> | ||
EcAMP-1 is an antimicrobial peptide of 37 amino acids | EcAMP-1 is an antimicrobial peptide of 37 amino acids | ||
| Line 405: | Line 422: | ||
<p class="title4">Design</p> | <p class="title4">Design</p> | ||
<p class="texte">This part is composed of the constitutive promoter P<sub>veg</sub> and | <p class="texte">This part is composed of the constitutive promoter P<sub>veg</sub> and | ||
| - | strong RBS for <i> | + | strong RBS for <i>Bacillus subtilis</i> |
(<a href="http://parts.igem.org/Part:BBa_K733013">K733013</a>) and | (<a href="http://parts.igem.org/Part:BBa_K733013">K733013</a>) and | ||
the open reading frame of EcAMP-1.<br> | the open reading frame of EcAMP-1.<br> | ||
| Line 418: | Line 435: | ||
<p class="texte">Svetlana B. Nolde, Alexander A. Vassilevski, Eugene A. Rogozhin, Nikolay A. Barinov, Tamara A. Balashova, Olga V. Samsonova, Yuri V. Baranov, Alexey S. Arseniev and Eugene V. Grishin. <b>Disulfide-stabilized Helical Hairpin Structure and Activity of a Novel Antifungal Peptide EcAMP1 from Seeds of Barnyard Grass (Echinochloa crus-galli)</b>. The journal of Biological Chemistry. 2011, Vol. 286, 25145–25153</p> | <p class="texte">Svetlana B. Nolde, Alexander A. Vassilevski, Eugene A. Rogozhin, Nikolay A. Barinov, Tamara A. Balashova, Olga V. Samsonova, Yuri V. Baranov, Alexey S. Arseniev and Eugene V. Grishin. <b>Disulfide-stabilized Helical Hairpin Structure and Activity of a Novel Antifungal Peptide EcAMP1 from Seeds of Barnyard Grass (Echinochloa crus-galli)</b>. The journal of Biological Chemistry. 2011, Vol. 286, 25145–25153</p> | ||
| - | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone(' | + | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 8); return false">Collapse</a></p> |
| - | </div> | + | </div></td></table> |
| - | + | <table> | |
| - | < | + | <td valign="top"; align="center";> |
| - | <div class=" | + | <p class="title2"; "line-height:6px"; "padding:10px";> |
| + | <a href="http://parts.igem.org/Part:BBa_K1364011">BBa_K1364011</a></p></td> | ||
| + | <td valign="top"; align="center";> | ||
| + | <div class="technology2"> P<sub>veg</sub> - SpoVG - Antifungal EcAMP-1 - Double terminator</div> | ||
| + | <div class="thelanguage2"> | ||
<p class="texte" id="select10"> | <p class="texte" id="select10"> | ||
| Line 434: | Line 455: | ||
<p class="title4">Design</p> | <p class="title4">Design</p> | ||
<p class="texte">This part is composed of the constitutive promoter P<sub>veg</sub> and | <p class="texte">This part is composed of the constitutive promoter P<sub>veg</sub> and | ||
| - | strong RBS for <i> | + | strong RBS for <i>Bacillus subtilis</i> |
(<a href="http://parts.igem.org/Part:BBa_K733013">K733013</a>), | (<a href="http://parts.igem.org/Part:BBa_K733013">K733013</a>), | ||
the open reading frame of EcAMP-1 and | the open reading frame of EcAMP-1 and | ||
a double terminator (<a href="http://parts.igem.org/Part:BBa_B0015">B0015</a>).<br> | a double terminator (<a href="http://parts.igem.org/Part:BBa_B0015">B0015</a>).<br> | ||
| - | The EcAMP-1 part was codon optimized for <i> | + | The EcAMP-1 part was codon optimized for <i>Escherichia coli</i> by the iGEM Utah State team |
and thanks to the Life Technologies GeneArt software program. </p> | and thanks to the Life Technologies GeneArt software program. </p> | ||
<img src="https://static.igem.org/mediawiki/2014/7/74/BBa_K1364011.png"> | <img src="https://static.igem.org/mediawiki/2014/7/74/BBa_K1364011.png"> | ||
| Line 448: | Line 469: | ||
<p class="texte">Svetlana B. Nolde, Alexander A. Vassilevski, Eugene A. Rogozhin, Nikolay A. Barinov, Tamara A. Balashova, Olga V. Samsonova, Yuri V. Baranov, Alexey S. Arseniev and Eugene V. Grishin. <b>Disulfide-stabilized Helical Hairpin Structure and Activity of a Novel Antifungal Peptide EcAMP1 from Seeds of Barnyard Grass (Echinochloa crus-galli)</b>. The journal of Biological Chemistry. 2011, Vol. 286, 25145–25153</p> | <p class="texte">Svetlana B. Nolde, Alexander A. Vassilevski, Eugene A. Rogozhin, Nikolay A. Barinov, Tamara A. Balashova, Olga V. Samsonova, Yuri V. Baranov, Alexey S. Arseniev and Eugene V. Grishin. <b>Disulfide-stabilized Helical Hairpin Structure and Activity of a Novel Antifungal Peptide EcAMP1 from Seeds of Barnyard Grass (Echinochloa crus-galli)</b>. The journal of Biological Chemistry. 2011, Vol. 286, 25145–25153</p> | ||
| - | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone(' | + | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 9); return false">Collapse</a></p> |
| - | </div> | + | </div></td></table> |
| - | + | <table> | |
| - | < | + | <td valign="top"; align="center";> |
| - | <div class=" | + | <p class="title2"; "line-height:6px"; "padding:10px";> |
| + | <a href="http://parts.igem.org/Part:BBa_K1364019">BBa_K1364019</a></p></td> | ||
| + | <td valign="top"; align="center";> | ||
| + | <div class="technology2">P<sub>veg</sub> - RBS - Antifungal EcAMP-1 (revised with a stop codon) - Double terminator</div> | ||
| + | <div class="thelanguage2"> | ||
<p class="texte">EcAMP-1 is an antimicrobial peptide of 37 amino acids | <p class="texte">EcAMP-1 is an antimicrobial peptide of 37 amino acids | ||
| Line 465: | Line 490: | ||
<p class="title4">Design</p> | <p class="title4">Design</p> | ||
<p class="texte">This part is composed of the constitutive promoter P<sub>veg</sub> and | <p class="texte">This part is composed of the constitutive promoter P<sub>veg</sub> and | ||
| - | strong RBS for <i> | + | strong RBS for <i>Bacillus subtilis</i> |
(<a href="http://parts.igem.org/Part:BBa_K733013">K733013</a>), | (<a href="http://parts.igem.org/Part:BBa_K733013">K733013</a>), | ||
<b>the revised open reading frame of EcAMP-1 (with a stop codon) </b> and | <b>the revised open reading frame of EcAMP-1 (with a stop codon) </b> and | ||
a double terminator (<a href="http://parts.igem.org/Part:BBa_B0015">B0015</a>).<br> | a double terminator (<a href="http://parts.igem.org/Part:BBa_B0015">B0015</a>).<br> | ||
| - | The EcAMP-1 part was codon optimized for <i> | + | The EcAMP-1 part was codon optimized for <i>Escherichia coli</i> by the iGEM Utah State team |
and thanks to the Life Technologies GeneArt software program. </p> | and thanks to the Life Technologies GeneArt software program. </p> | ||
<img src="https://static.igem.org/mediawiki/2014/7/74/BBa_K1364011.png"> | <img src="https://static.igem.org/mediawiki/2014/7/74/BBa_K1364011.png"> | ||
| Line 479: | Line 504: | ||
<p class="texte">Svetlana B. Nolde, Alexander A. Vassilevski, Eugene A. Rogozhin, Nikolay A. Barinov, Tamara A. Balashova, Olga V. Samsonova, Yuri V. Baranov, Alexey S. Arseniev and Eugene V. Grishin. <b>Disulfide-stabilized Helical Hairpin Structure and Activity of a Novel Antifungal Peptide EcAMP1 from Seeds of Barnyard Grass (Echinochloa crus-galli)</b>. The journal of Biological Chemistry. 2011, Vol. 286, 25145–25153</p> | <p class="texte">Svetlana B. Nolde, Alexander A. Vassilevski, Eugene A. Rogozhin, Nikolay A. Barinov, Tamara A. Balashova, Olga V. Samsonova, Yuri V. Baranov, Alexey S. Arseniev and Eugene V. Grishin. <b>Disulfide-stabilized Helical Hairpin Structure and Activity of a Novel Antifungal Peptide EcAMP1 from Seeds of Barnyard Grass (Echinochloa crus-galli)</b>. The journal of Biological Chemistry. 2011, Vol. 286, 25145–25153</p> | ||
| - | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone(' | + | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 13); return false">Collapse</a></p> |
| - | </div> | + | </div></td></table> |
<br></br> | <br></br> | ||
| Line 486: | Line 511: | ||
<p class="title3">Fungicide operons</p> | <p class="title3">Fungicide operons</p> | ||
| - | < | + | <table> |
| - | <div class=" | + | <td valign="top"; align="center";> |
| - | + | <p class="title2"; "line-height:6px"; "padding:10px";> | |
| + | <a href="http://parts.igem.org/Part:BBa_K1364013">BBa_K1364013</a></p></td> | ||
| + | <td valign="top"; align="center";> | ||
| + | <div class="technology2">P<sub>veg</sub> - SpoVG - Antifungals GAFP-1 and D4E1 - Double terminator</div> | ||
| + | <div class="thelanguage2"> | ||
<p class="texte"> | <p class="texte"> | ||
<p class="title4">Design</p> | <p class="title4">Design</p> | ||
<p class="texte">This part is designed for the co-expression of two different peptides | <p class="texte">This part is designed for the co-expression of two different peptides | ||
with anti-fungal activities : D4E1 and GAFP-1. | with anti-fungal activities : D4E1 and GAFP-1. | ||
| - | It is composed of the strong, constitutive promoter of <i> | + | It is composed of the strong, constitutive promoter of <i>Bacillus subtilis</i> |
| - | P<sub>veg</sub> (<a href="http://parts.igem.org/wiki/index.php/Part:BBa_K823003"> | + | P<sub>veg</sub> (<a href="http://parts.igem.org/wiki/index.php/Part:BBa_K823003">BBa_K823003</a>) |
and the translation unit with GAFP-1 and D4E1 (BBa_K1364012).</p> | and the translation unit with GAFP-1 and D4E1 (BBa_K1364012).</p> | ||
<img src="https://static.igem.org/mediawiki/2014/4/40/BBa_K1364013.png"> | <img src="https://static.igem.org/mediawiki/2014/4/40/BBa_K1364013.png"> | ||
| Line 504: | Line 533: | ||
See <a href="http://parts.igem.org/Part:BBa_K1364002">BBa_K1364002</a> and <a href="http://parts.igem.org/Part:BBa_K1364003">BBa_K1364003</a>.</p> | See <a href="http://parts.igem.org/Part:BBa_K1364002">BBa_K1364002</a> and <a href="http://parts.igem.org/Part:BBa_K1364003">BBa_K1364003</a>.</p> | ||
| - | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone(' | + | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 10); return false">Collapse</a></p> |
| - | </div> | + | </div></td></table> |
<br></br> | <br></br> | ||
| Line 511: | Line 540: | ||
<p class="title2">Basic tools</p> | <p class="title2">Basic tools</p> | ||
| - | < | + | <table> |
| - | <div class=" | + | <td valign="top"; align="center";> |
| + | <p class="title2"; "line-height:6px"; "padding:10px";> | ||
| + | <a href="http://parts.igem.org/Part:BBa_K1364015">BBa_K1364015</a></p></td> | ||
| + | <td valign="top"; align="center";> | ||
| + | <div class="technology2">P<sub>veg</sub> + RFP</div> | ||
| + | <div class="thelanguage2"> | ||
<p class="texte"><!--Compléter--> | <p class="texte"><!--Compléter--> | ||
| - | This part is designed to enable the expression of a Red Fluorescent Protein in Bacillus subtilis under the control of a constitutive promoter | + | This part is designed to enable the expression of a Red Fluorescent Protein in <i>Bacillus subtilis</i> under the control of a constitutive promoter P<sub>veg</sub>. This construction has been checked by sequencing and has shown to work also in <i>Escherichia coli</i> |
</p> | </p> | ||
<img src="https://static.igem.org/mediawiki/parts/6/69/Pveg%2BRFP.jpg"> | <img src="https://static.igem.org/mediawiki/parts/6/69/Pveg%2BRFP.jpg"> | ||
<p class="title4">Design</p> | <p class="title4">Design</p> | ||
| - | <p class="texte">This part is composed of a constitutive promoter | + | <p class="texte">This part is composed of a constitutive promoter P<sub>veg</sub> (BBa_K823003), spoVG RBS (BBa_KK143021), the coding sequence of the RFP (E1010 from BBa_K606013) and a double terminator (BBa_KB0015) </p> |
<p class="title4">Type</p> | <p class="title4">Type</p> | ||
<p class="texte">Reporter</p> | <p class="texte">Reporter</p> | ||
<p class="title4">Tests</p> | <p class="title4">Tests</p> | ||
| - | <p class="texte">This part was tested in both E.coli and B. subtilis. The sequences was verified by sequencing. </p> | + | <p class="texte">This part was tested in both <i>E.coli</i> and <i>B. subtilis</i>. The sequences was verified by sequencing. </p> |
| - | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone(' | + | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 10); return false">Collapse</a></p> |
| - | </div> | + | </div></td></table> |
| - | < | + | <table> |
| - | <div class=" | + | <td valign="top"; align="center";> |
| + | <p class="title2"; "line-height:6px"; "padding:10px";> | ||
| + | <a href="http://parts.igem.org/Part:BBa_K1364016">BBa_K1364016</a></p></td> | ||
| + | <td valign="top"; align="center";> | ||
| + | <div class="technology2"> P<sub>lepA</sub> + RFP</div> | ||
| + | <div class="thelanguage2"> | ||
<p class="texte"><!--Compléter--> | <p class="texte"><!--Compléter--> | ||
| - | This part is designed to enable the expression of a Red Fluorescent Protein in Bacillus subtilis under the control of a constitutive promoter | + | This part is designed to enable the expression of a Red Fluorescent Protein in <i>Bacillus subtilis</i> under the control of a constitutive promoter P<sub>lepA</sub>. This construction has been checked by sequencing and has shown to work also in <i>Escherichia coli</i> |
</p> | </p> | ||
<img src="https://static.igem.org/mediawiki/parts/1/12/PlepA%2BRFP.jpg"> | <img src="https://static.igem.org/mediawiki/parts/1/12/PlepA%2BRFP.jpg"> | ||
<p class="title4">Design</p> | <p class="title4">Design</p> | ||
| - | <p class="texte">This part is composed of a constitutive promoter | + | <p class="texte">This part is composed of a constitutive promoter P<sub>lepA</sub> (BBa_K823002), spoVG RBS (BBa_K143021), the coding sequence of the RFP (E1010 from BBa_K606013) and a double terminator (BBa_B0015) </p> |
<p class="title4">Type</p> | <p class="title4">Type</p> | ||
<p class="texte">Reporter</p> | <p class="texte">Reporter</p> | ||
<p class="title4">Tests</p> | <p class="title4">Tests</p> | ||
| - | <p class="texte">This part was tested in both E.coli and B. subtilis. The sequences was verified by sequencing.</p> | + | <p class="texte">This part was tested in both <i>E.coli</i> and <i>B. subtilis</i>. The sequences was verified by sequencing.</p> |
| - | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone(' | + | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 11); return false">Collapse</a></p> |
| - | </div> | + | </div></td></table> |
| - | < | + | <table> |
| - | <div class=" | + | <td valign="top"; align="center";> |
| + | <p class="title2"; "line-height:6px"; "padding:10px";> | ||
| + | <a href="http://parts.igem.org/Part:BBa_K1364017">BBa_K1364017</a></p></td> | ||
| + | <td valign="top"; align="center";> | ||
| + | <div class="technology2">P<sub>lepA</sub> + RBS SpoVG</div> | ||
| + | <div class="thelanguage2"> | ||
<p class="texte"><!--Compléter--> | <p class="texte"><!--Compléter--> | ||
| - | + | P<sub>lepA</sub> is a constitutive promoter in <i>Bacillus subtilis</i> (BBa_K823002) coupled with a RBS spoVG (BBa_K143021). To get the highest level of translation from this Promoter-RBS combination it must be connected to a coding region preceded by a coding region prefix. A standard prefix will increase the distance between the RBS and the start codon, reducing translational efficiency. | |
| - | This construction is also working with | + | This construction is also working with <i>Escherichia coli</i> and has been verified by sequencing. |
</p> | </p> | ||
<img src="https://static.igem.org/mediawiki/parts/d/da/PlepA%2BSpoVG.jpg"> | <img src="https://static.igem.org/mediawiki/parts/d/da/PlepA%2BSpoVG.jpg"> | ||
| Line 561: | Line 605: | ||
<p class="texte">This part was checked by sequencing</p> | <p class="texte">This part was checked by sequencing</p> | ||
| + | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 12); return false">Collapse</a></p> | ||
| + | </div></td></table> | ||
| - | < | + | <table> |
| - | </div> | + | <td valign="top"; align="center";> |
| + | <p class="title2"; "line-height:6px"; "padding:10px";> | ||
| + | <a href="http://parts.igem.org/Part:BBa_K1364021">BBa_K1364021</a></p></td> | ||
| + | <td valign="top"; align="center";> | ||
| + | <div class="technology2">Integrative plasmid for <i>Bacillus subtilis</i> (pSB<sub>bs</sub>4E)</div> | ||
| + | <div class="thelanguage2"> | ||
| - | + | <p class="texte">This part is an empty backbone vector for the usage in <i>Bacillus subtilis</i>. It integrates in the thrC locus and can be selected with Erythromycin . It has an Ampicillin resistance for cloning in <i>Escherichia coli</i>. The backbones contains an RFP in the BioBrick site (BBa_KJ04450) to facilitate the cloning in <i>E.coli</i>. <br> | |
| - | + | ||
| - | + | ||
| - | + | ||
| - | <p class="texte">This part is an empty backbone vector for the usage in <i>Bacillus subtilis</i>. It integrates in the thrC locus and can be selected with Erythromycin . It has an Ampicillin resistance for cloning in | + | |
The handling of this type of vector is described <a href="https://2014.igem.org/Team:Toulouse/Notebook/Protocols#select6">here</a>. | The handling of this type of vector is described <a href="https://2014.igem.org/Team:Toulouse/Notebook/Protocols#select6">here</a>. | ||
| Line 581: | Line 628: | ||
<p class="title4">References</p> | <p class="title4">References</p> | ||
<p class="texte"> | <p class="texte"> | ||
| - | R. Bernard, K.A. Marquis, and D.Z. Rudner. <b>Nucleoid occlusion prevents cell division during replication fork arrest in Bacillus subtilis</b>Mol Microbiol. Nov 2010; 78(4): 866–882.</p> | + | R. Bernard, K.A. Marquis, and D.Z. Rudner. <b>Nucleoid occlusion prevents cell division during replication fork arrest in <i>Bacillus subtilis</i></b>Mol Microbiol. Nov 2010; 78(4): 866–882.</p> |
| - | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone(' | + | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 13); return false">Collapse</a></p> |
| - | </div> | + | </div></td></table> |
<br></br> | <br></br> | ||
Revision as of 18:06, 17 October 2014
Parts
What did we send to the Registry?
Results > Parts > Submitted parts
We deposited 16 new BioBrick parts in the Registry.
Most of them were cloned into the standard plasmid pSB1C3, tested and sequenced.
Chemotaxis
|
N-acetylglucosamine based chemotaxis for Bacillus subtilis
This part is designed to enable a N-acetylglucosamine based chemotaxis in B. subtilis. Design The encoded protein is a chimera of two proteins: 
Type Translation unit Tests This part is not tested yet. References Uniprot: |
|
Pveg + N-acetylglucosamine based chemotaxis for Bacillus subtilis
This part is designed to enable a N-acetylglucosamine based chemotaxis in Bacillus subtilis. Design This expression cassette is designed for the expression of an antifungal peptide, D4E1 and for its secretion in B. subtilis. It is composed of the strong, constitutive promoter of B. subtilis Pveg (K823003), strong RBS for B. subtilis (K780002), the open reading frame of D4E1 and a double terminator (B0015) 
Type Generator Tests This part was not tested yet. References Uniprot: |
Binding
|
Pveg + Chitin Binding Cell Wall protein
This part is designed to enable the binding of Bacillus subtilis to the fungi wall made of chitin. Design This part is composed of a Cell Wall Binding (CWB) sequence
and a Chitin Binding Domain (CBD) linked with a 6 amino acids linker. 
Type Generator Tests This part was tested with chitin beads. (See Binding module) References - Wong E, Vaaje-Kolstad G, Ghosh A, Hurtado-Guerrero R, Konarev PV, et al. (2012) |
Fungicides
GAFP-1
|
RBS - Antifungal GAFP-1
The Gastrodia anti-fungal protein(GAFP-1), also known as gastrodianin, is a mannose and chitin binding lectin originating from the Asiatic orchid Gastrodia elata, a traditional Chinese medicinal herb cultured for thousands of years. GAFP-1 is composed of 15 amino acids LDSLSFSYNNFEEDD and is able to inhibit the growth of multiples species of plant pathogenic fungi. Design This part is composed of a Strong RBS (K780002) and the open reading frame of the Gastrodia anti-fungal protein 1 (GAFP-1) optimized for its expression and its secretion in Bacillus subtilis. The codon optimization was made thanks to the DNA 2.0 software program. 
Type Translational unit Tests This part was tested on the fungi Trichoderma reesei (See Fungicides module) References - Wong E, Vaaje-Kolstad G, Ghosh A, Hurtado-Guerrero R, Konarev PV, et al. (2012) |
|
RBS + Antifungal GAFP-1 + Double terminator
The Gastrodia anti-fungal protein(GAFP-1), also known as gastrodianin, is a mannose and chitin binding lectin originating from the Asiatic orchid Gastrodia elata, a traditional Chinese medicinal herb cultured for thousands of years. GAFP-1 is composed of 15 amino acids LDSLSFSYNNFEEDD and is able to inhibit the growth of multiples species of plant pathogenic fungi. Design This part is composed of a Strong RBS (K780002), the open reading frame of the Gastrodia anti-fungal protein 1 (GAFP-1) and a double terminator B0015. optimized for its expression and its secretion in Bacillus subtilis. The codon optimization was made thanks to the DNA 2.0 software program. 
Type Composite part Tests This part was tested on the fungi Trichoderma reesei (See Fungicides module) References - Wong E, Vaaje-Kolstad G, Ghosh A, Hurtado-Guerrero R, Konarev PV, et al. (2012) |
|
Pveg - strong RBS - Antifungal GAFP-1 - Double terminator
This expression cassette is designed for the expression and secretion of the Gastrodia anti-fungal protein(GAFP-1), also known as gastrodianin. GAFP-1 is a mannose and chitin binding lectin originating from the Asiatic orchid Gastrodia elata, a traditional Chinese medicinal herb cultured for thousands of years. GAFP-1 is composed of 15 amino acids LDSLSFSYNNFEEDD and is able to inhibit the growth of multiples species of plant pathogenic fungi. Design This part is composed of the strong, constitutive promoter of Bacillus subtilis Pveg (K823003), strong RBS (K780002), the open reading frame of the Gastrodia anti-fungal protein 1 (GAFP-1) and a double terminator (B0015). optimized for its expression and its secretion in B. subtilis. The codon optimization was made thanks to the DNA 2.0 software program. 
Type Generator Tests This part was tested on the fungi Trichoderma reesei (See Fungicides module) References - Wong E, Vaaje-Kolstad G, Ghosh A, Hurtado-Guerrero R, Konarev PV, et al. (2012) |
D4E1
|
RBS - Antifungal D4E1 - Double terminator
D4E1 is a linear synthetic peptide of 17 amino acids which has shown to have antifungal activities by complexing with a sterol present in conodial wall of a varety of fungi. Design This part is composed of a Strong RBS (K780002), the open reading frame of D4E1 and a Double terminator (B0015) optimized for its expression and its secretion in Bacillus subtilis. This part was optimized for the expression and its secretion in B. subtilis thanks to the DNA 2.0 software program. 
Type Translational unit Tests This part was tested on the fungi Trichoderma reesei (See Fungicides module) References - De Lucca AJ, Bland JM, Grimm C, Jacks TJ, Cary JW, Jaynes JM, Cleveland TE, Walsh TJ. Fungicidal properties, sterol binding, and proteolytic resistance of the synthetic peptide D4E1. |
|
Pveg - RBS - Antifungal D4E1 - Double Terminator
D4E1 is a linear synthetic peptide of 17 amino acids which has shown to have antifungal activities by complexing with a sterol present in conodial wall of a varety of fungi. Design This part is composed of the strong, constitutive promoter of Bacillus subtilis Pveg (K823003), a strong RBS (K780002), the open reading frame of D4E1 and a double terminator (B0015). This part was optimized for the expression and its secretion in B.subtilis thanks to the DNA 2.0 software program. 
Type Translational unit Tests This part was tested on the fungi Trichoderma reesei (See Fungicides module) References - De Lucca AJ, Bland JM, Grimm C, Jacks TJ, Cary JW, Jaynes JM, Cleveland TE, Walsh TJ. Fungicidal properties, sterol binding, and proteolytic resistance of the synthetic peptide D4E1. |
EcAMP-1
|
Pveg - SpoVG - Antifungal EcAMP-1
EcAMP-1 is an antimicrobial peptide of 37 amino acids
originated from the specie Echinochloa crus-galli,
a type of wild grass.
This peptide has a particular structure : it is helical because of two disulfide bonds. Design This part is composed of the constitutive promoter Pveg and
strong RBS for Bacillus subtilis
(K733013) and
the open reading frame of EcAMP-1. 
Type Generator Tests This part is not tested yet. References Svetlana B. Nolde, Alexander A. Vassilevski, Eugene A. Rogozhin, Nikolay A. Barinov, Tamara A. Balashova, Olga V. Samsonova, Yuri V. Baranov, Alexey S. Arseniev and Eugene V. Grishin. Disulfide-stabilized Helical Hairpin Structure and Activity of a Novel Antifungal Peptide EcAMP1 from Seeds of Barnyard Grass (Echinochloa crus-galli). The journal of Biological Chemistry. 2011, Vol. 286, 25145–25153 |
|
Pveg - SpoVG - Antifungal EcAMP-1 - Double terminator
EcAMP-1 is an antimicrobial peptide of 37 amino acids
originated from the specie Echinochloa crus-galli,
a type of wild grass.
This peptide has a particular structure : it is helical because of two disulfide bonds. Design This part is composed of the constitutive promoter Pveg and
strong RBS for Bacillus subtilis
(K733013),
the open reading frame of EcAMP-1 and
a double terminator (B0015). 
Type Generator Tests This part is not tested yet. References Svetlana B. Nolde, Alexander A. Vassilevski, Eugene A. Rogozhin, Nikolay A. Barinov, Tamara A. Balashova, Olga V. Samsonova, Yuri V. Baranov, Alexey S. Arseniev and Eugene V. Grishin. Disulfide-stabilized Helical Hairpin Structure and Activity of a Novel Antifungal Peptide EcAMP1 from Seeds of Barnyard Grass (Echinochloa crus-galli). The journal of Biological Chemistry. 2011, Vol. 286, 25145–25153 |
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Pveg - RBS - Antifungal EcAMP-1 (revised with a stop codon) - Double terminator
EcAMP-1 is an antimicrobial peptide of 37 amino acids
originated from the specie Echinochloa crus-galli,
a type of wild grass.
This peptide has a particular structure : it is helical because of two disulfide bonds. Design This part is composed of the constitutive promoter Pveg and
strong RBS for Bacillus subtilis
(K733013),
the revised open reading frame of EcAMP-1 (with a stop codon) and
a double terminator (B0015).
Type Composite Tests This part was not tested yet References Svetlana B. Nolde, Alexander A. Vassilevski, Eugene A. Rogozhin, Nikolay A. Barinov, Tamara A. Balashova, Olga V. Samsonova, Yuri V. Baranov, Alexey S. Arseniev and Eugene V. Grishin. Disulfide-stabilized Helical Hairpin Structure and Activity of a Novel Antifungal Peptide EcAMP1 from Seeds of Barnyard Grass (Echinochloa crus-galli). The journal of Biological Chemistry. 2011, Vol. 286, 25145–25153 |
Fungicide operons
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Pveg - SpoVG - Antifungals GAFP-1 and D4E1 - Double terminator
Design This part is designed for the co-expression of two different peptides with anti-fungal activities : D4E1 and GAFP-1. It is composed of the strong, constitutive promoter of Bacillus subtilis Pveg (BBa_K823003) and the translation unit with GAFP-1 and D4E1 (BBa_K1364012). 
Type Generator Tests This part was tested on the fungi Trichoderma reesei (See Fungicides module) References See BBa_K1364002 and BBa_K1364003. |
Basic tools
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Pveg + RFP
This part is designed to enable the expression of a Red Fluorescent Protein in Bacillus subtilis under the control of a constitutive promoter Pveg. This construction has been checked by sequencing and has shown to work also in Escherichia coli 
Design This part is composed of a constitutive promoter Pveg (BBa_K823003), spoVG RBS (BBa_KK143021), the coding sequence of the RFP (E1010 from BBa_K606013) and a double terminator (BBa_KB0015) Type Reporter Tests This part was tested in both E.coli and B. subtilis. The sequences was verified by sequencing. |
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PlepA + RFP
This part is designed to enable the expression of a Red Fluorescent Protein in Bacillus subtilis under the control of a constitutive promoter PlepA. This construction has been checked by sequencing and has shown to work also in Escherichia coli 
Design This part is composed of a constitutive promoter PlepA (BBa_K823002), spoVG RBS (BBa_K143021), the coding sequence of the RFP (E1010 from BBa_K606013) and a double terminator (BBa_B0015) Type Reporter Tests This part was tested in both E.coli and B. subtilis. The sequences was verified by sequencing. |
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PlepA + RBS SpoVG
PlepA is a constitutive promoter in Bacillus subtilis (BBa_K823002) coupled with a RBS spoVG (BBa_K143021). To get the highest level of translation from this Promoter-RBS combination it must be connected to a coding region preceded by a coding region prefix. A standard prefix will increase the distance between the RBS and the start codon, reducing translational efficiency. This construction is also working with Escherichia coli and has been verified by sequencing. 
Type Composite Tests This part was checked by sequencing |
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Integrative plasmid for Bacillus subtilis (pSBbs4E)
This part is an empty backbone vector for the usage in Bacillus subtilis. It integrates in the thrC locus and can be selected with Erythromycin . It has an Ampicillin resistance for cloning in Escherichia coli. The backbones contains an RFP in the BioBrick site (BBa_KJ04450) to facilitate the cloning in E.coli. Design We digested pKL190 plasmid and PCR products of BBa_J04450 with BamHI and EcoRI. We proceded to ligation. Type Backbone Tests This part was tested by transforming B. subtilis. Clones obtained were Threonine-dependant and resistant to Erythromycin (10µg/ml). References R. Bernard, K.A. Marquis, and D.Z. Rudner. Nucleoid occlusion prevents cell division during replication fork arrest in Bacillus subtilisMol Microbiol. Nov 2010; 78(4): 866–882. |
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