"
Team:Toulouse/Result/parts/Submitted parts
From 2014.igem.org
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<a href="http://parts.igem.org/Part:BBa_K1364004">BBa_K1364004</a></p></td> | <a href="http://parts.igem.org/Part:BBa_K1364004">BBa_K1364004</a></p></td> | ||
<td valign="top"; align="center";> | <td valign="top"; align="center";> | ||
| - | <div class="technology2">P<sub>veg</sub> | + | <div class="technology2">P<sub>veg</sub> - N-acetylglucosamine based chemotaxis for <i>Bacillus subtilis</i></div> |
<div class="thelanguage2"> | <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> | ||
| Line 200: | Line 200: | ||
D4E1 and for its secretion in <i>B. subtilis</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"> | + | P<sub>veg</sub> (<a href="http://parts.igem.org/wiki/index.php/Part:BBa_K823003">BBa_K823003</a>), |
| - | strong RBS for <i>B. subtilis</i> (<a href="http://parts.igem.org/Part:BBa_K780002"> | + | strong RBS for <i>B. subtilis</i> (<a href="http://parts.igem.org/Part:BBa_K780002">BBa_K780002</a>), |
the open reading frame of D4E1 | the open reading frame of D4E1 | ||
| - | and a double terminator (<a href="http://parts.igem.org/Part:BBa_B0015"> | + | and a double terminator (<a href="http://parts.igem.org/Part:BBa_B0015">BBa_B0015</a>)</p> |
<img src="https://static.igem.org/mediawiki/2014/3/35/BBa_K1364004.png"> | <img src="https://static.igem.org/mediawiki/2014/3/35/BBa_K1364004.png"> | ||
<p class="title4">Type</p> | <p class="title4">Type</p> | ||
| Line 225: | Line 225: | ||
<a href="http://parts.igem.org/Part:BBa_K1364005">BBa_K1364005</a></p></td> | <a href="http://parts.igem.org/Part:BBa_K1364005">BBa_K1364005</a></p></td> | ||
<td valign="top"; align="center";> | <td valign="top"; align="center";> | ||
| - | <div class="technology2"></a>P<sub>veg</sub> | + | <div class="technology2"></a>P<sub>veg</sub> - Chitin Binding Cell Wall protein</div> |
<div class="thelanguage2"> | <div class="thelanguage2"> | ||
| Line 244: | Line 244: | ||
<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 <i>Vibrio cholerae</i> 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> | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 2); return false">Collapse</a></p> | ||
| Line 267: | Line 267: | ||
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> | ||
| - | <p class="texte">This part is composed of a Strong RBS (<a href="http://parts.igem.org/Part:BBa_K780002"> | + | <p class="texte">This part is composed of a Strong RBS (<a href="http://parts.igem.org/Part:BBa_K780002">BBa_K780002</a>) and |
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) | ||
optimized for its expression and its secretion in <i>Bacillus subtilis</i>. | optimized for its expression and its secretion in <i>Bacillus subtilis</i>. | ||
| Line 278: | Line 278: | ||
<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</p> | + | - The <i>Vibrio cholerae</i> 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> | <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> | </div></td></table> | ||
| Line 287: | Line 287: | ||
<a href="http://parts.igem.org/Part:BBa_K1364007">BBa_K1364007</a></p></td> | <a href="http://parts.igem.org/Part:BBa_K1364007">BBa_K1364007</a></p></td> | ||
<td valign="top"; align="center";> | <td valign="top"; align="center";> | ||
| - | <div class="technology2">RBS | + | <div class="technology2">RBS - Antifungal GAFP-1 + Double terminator</div> |
<div class="thelanguage2"> | <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 | ||
| Line 293: | Line 293: | ||
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> | ||
| - | <p class="texte">This part is composed of a Strong RBS (<a href="http://parts.igem.org/Part:BBa_K780002"> | + | <p class="texte">This part is composed of a Strong RBS (<a href="http://parts.igem.org/Part:BBa_K780002">BBa_K780002</a>), |
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"> | + | and a double terminator <a href="http://parts.igem.org/Part:BBa_B0015">BBa_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>. | ||
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> | ||
| Line 305: | 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</p> | + | - The <i>Vibrio cholerae</i> 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> | <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> | </div></td></table> | ||
| Line 320: | Line 320: | ||
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> | ||
| - | <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"> | + | <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">BBa_K823003</a>), |
| - | strong RBS (<a href="http://parts.igem.org/Part:BBa_K780002"> | + | strong RBS (<a href="http://parts.igem.org/Part:BBa_K780002">BBa_K780002</a>), |
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>). | ||
| Line 333: | 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 <i>Vibrio cholerae</i> 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('technology2', 5); return false">Collapse</a></p> | ||
| Line 354: | Line 354: | ||
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 a Strong RBS (<a href="http://parts.igem.org/Part:BBa_K780002"> | + | <p class="texte">This part is composed of a Strong RBS (<a href="http://parts.igem.org/Part:BBa_K780002">BBa_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"> | + | a Double terminator (<a href="http://parts.igem.org/Part:BBa_B0015">BBa_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>B. subtilis </i> | This part was optimized for the expression and its secretion in <i>B. subtilis </i> | ||
| Line 384: | Line 384: | ||
<p class="title4">Design</p> | <p class="title4">Design</p> | ||
<p class="texte">This part is composed of the strong, constitutive promoter of <i>Bacillus subtilis</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"> | + | P<sub>veg</sub> (<a href="http://parts.igem.org/wiki/index.php/Part:BBa_K823003">BBa_K823003</a>), |
| - | a strong RBS (<a href="http://parts.igem.org/Part:BBa_K780002"> | + | a strong RBS (<a href="http://parts.igem.org/Part:BBa_K780002">BBa_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"> | + | a double terminator (<a href="http://parts.igem.org/Part:BBa_B0015">BBa_B0015</a>). |
This part was optimized for the expression and its secretion in <i>B.subtilis </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> | ||
| Line 423: | Line 423: | ||
<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>Bacillus subtilis</i> | strong RBS for <i>Bacillus subtilis</i> | ||
| - | (<a href="http://parts.igem.org/Part:BBa_K733013"> | + | (<a href="http://parts.igem.org/Part:BBa_K733013">BBa_K733013</a>) and |
the open reading frame of EcAMP-1.<br> | the open reading frame of EcAMP-1.<br> | ||
The EcAMP-1 part was codon optimized for <i>E. coli</i> by the iGEM Utah State team | The EcAMP-1 part was codon optimized for <i>E. coli</i> by the iGEM Utah State team | ||
| Line 433: | Line 433: | ||
<p class="texte">This part is not tested yet.</p> | <p class="texte">This part is not tested yet.</p> | ||
<p class="title4">References</p> | <p class="title4">References</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 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 (<i>Echinochloa crus-galli</i>)</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('technology2', 8); return false">Collapse</a></p> | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 8); return false">Collapse</a></p> | ||
| Line 456: | Line 456: | ||
<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>Bacillus subtilis</i> | strong RBS for <i>Bacillus subtilis</i> | ||
| - | (<a href="http://parts.igem.org/Part:BBa_K733013"> | + | (<a href="http://parts.igem.org/Part:BBa_K733013">BBa_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"> | + | a double terminator (<a href="http://parts.igem.org/Part:BBa_B0015">BBa_B0015</a>).<br> |
The EcAMP-1 part was codon optimized for <i>Escherichia coli</i> by the iGEM Utah State team | 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> | ||
| Line 467: | Line 467: | ||
<p class="texte">This part is not tested yet.</p> | <p class="texte">This part is not tested yet.</p> | ||
<p class="title4">References</p> | <p class="title4">References</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 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 (<i>Echinochloa crus-galli</i>)</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('technology2', 9); return false">Collapse</a></p> | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 9); return false">Collapse</a></p> | ||
| Line 491: | Line 491: | ||
<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>Bacillus subtilis</i> | strong RBS for <i>Bacillus subtilis</i> | ||
| - | (<a href="http://parts.igem.org/Part:BBa_K733013"> | + | (<a href="http://parts.igem.org/Part:BBa_K733013">BBa_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"> | + | a double terminator (<a href="http://parts.igem.org/Part:BBa_B0015">BBa_B0015</a>).<br> |
The EcAMP-1 part was codon optimized for <i>Escherichia coli</i> by the iGEM Utah State team | 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> | ||
| Line 502: | Line 502: | ||
<p class="texte">This part was not tested yet</p> | <p class="texte">This part was not tested yet</p> | ||
<p class="title4">References</p> | <p class="title4">References</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 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 (<i>Echinochloa crus-galli</i>)</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('technology2', 13); return false">Collapse</a></p> | <p style="text-align:right;font-size:1.3em;"><a href="#" class="collapseLink" onClick="ddaccordion.collapseone('technology2', 13); return false">Collapse</a></p> | ||
| Line 545: | Line 545: | ||
<a href="http://parts.igem.org/Part:BBa_K1364015">BBa_K1364015</a></p></td> | <a href="http://parts.igem.org/Part:BBa_K1364015">BBa_K1364015</a></p></td> | ||
<td valign="top"; align="center";> | <td valign="top"; align="center";> | ||
| - | <div class="technology2">P<sub>veg</sub> | + | <div class="technology2">P<sub>veg</sub> - RFP</div> |
<div class="thelanguage2"> | <div class="thelanguage2"> | ||
| Line 557: | Line 557: | ||
<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 <i>E.coli</i> and <i>B. subtilis</i>. 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> |
| Line 568: | Line 568: | ||
<a href="http://parts.igem.org/Part:BBa_K1364016">BBa_K1364016</a></p></td> | <a href="http://parts.igem.org/Part:BBa_K1364016">BBa_K1364016</a></p></td> | ||
<td valign="top"; align="center";> | <td valign="top"; align="center";> | ||
| - | <div class="technology2"> P<sub>lepA</sub> | + | <div class="technology2"> P<sub>lepA</sub> - RFP</div> |
<div class="thelanguage2"> | <div class="thelanguage2"> | ||
| Line 581: | Line 581: | ||
<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 <i>E.coli</i> and <i>B. subtilis</i>. 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> |
| Line 592: | Line 592: | ||
<a href="http://parts.igem.org/Part:BBa_K1364017">BBa_K1364017</a></p></td> | <a href="http://parts.igem.org/Part:BBa_K1364017">BBa_K1364017</a></p></td> | ||
<td valign="top"; align="center";> | <td valign="top"; align="center";> | ||
| - | <div class="technology2">P<sub>lepA</sub> | + | <div class="technology2">P<sub>lepA</sub> - RBS SpoVG</div> |
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| - | <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 | + | <p class="texte">This part is an empty backbone vector for the usage in <i>Bacillus subtilis</i>. It is on biobrick version of pKL190 which integrates in the thrC locus and can be selected with Erythromycin . It has an Ampicillin resistance for cloning in <i>Escherichia coli</i>. The backbone contains an RFP in the BioBrick site (BBa_J04450) to facilitate the cloning in <i>E.coli</i>. <br> |
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>. | ||
Revision as of 18:21, 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
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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 (BBa_K823003), strong RBS for B. subtilis (BBa_K780002), the open reading frame of D4E1 and a double terminator (BBa_B0015) 
Type Generator Tests This part was not tested yet. References Uniprot: |
Binding
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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 (BBa_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 (BBa_K780002), the open reading frame of the Gastrodia anti-fungal protein 1 (GAFP-1) and a double terminator BBa_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 (BBa_K823003), strong RBS (BBa_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
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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 (BBa_K780002), the open reading frame of D4E1 and a Double terminator (BBa_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 (BBa_K823003), a strong RBS (BBa_K780002), the open reading frame of D4E1 and a double terminator (BBa_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
(BBa_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
(BBa_K733013),
the open reading frame of EcAMP-1 and
a double terminator (BBa_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 |
|
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
(BBa_K733013),
the revised open reading frame of EcAMP-1 (with a stop codon) and
a double terminator (BBa_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. |
|
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 |
|
Integrative plasmid for Bacillus subtilis (pSBbs4E)
This part is an empty backbone vector for the usage in Bacillus subtilis. It is on biobrick version of pKL190 which integrates in the thrC locus and can be selected with Erythromycin . It has an Ampicillin resistance for cloning in Escherichia coli. The backbone contains an RFP in the BioBrick site (BBa_J04450) 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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