Difference between revisions of "Team:Montpellier/WetLab Peptides"

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      <th class="references_title" colspan="2">References</th>
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      <td class="references_left">[1]</td>
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      <td class="references_right">Nguyen LT1, Haney EF, Vogel HJ. 2011. The expanding scope of antimicrobial peptide structures and their modes of action. <i>Trends Biotechnol</i> Vol 29, 464–472..</td>
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    </tr>
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      <td class="references_left">[2]</td>
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      <td class="references_right">Tanphaichitr, Nongnuj et al. 2018. Potential Use of Antimicrobial Peptides as Vaginal Spermicides/Microbicides. <i>Pharmaceuticals</i> 9.1 (2016): 13.</td>
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    </tr>
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      <td class="references_left">[3]</td>
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      <td class="references_right">Cavera, V. L., Volski, A., & Chikindas, M. L. 2015. The natural antimicrobial subtilosin A synergizes with lauramide arginine ethyl ester (LAE), ε-poly-l-lysine (polylysine), clindamycin phosphate and metronidazole, against the vaginal pathogen Gardnerella vaginalis. <i>Probiotics and antimicrobial proteins.</i> 7(2), 164-171.</td>
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    </tr>
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    <tr>
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      <td class="references_left">[4]</td>
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      <td class="references_right">Paul M. Himes, Scott E. Allen, Sungwon Hwang, and Albert A. Bowers. 2016.Production of Sactipeptides in Escherichia coli: Probing the Substrate Promiscuity of Subtilosin A Biosynthesis. <i>ACS Chemical Biology</i>. 11 (6), 1737-1744.</td>
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    </tr>
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      <td class="references_left">[5]</td>
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      <td class="references_right">Srinivas Suda et al., 2012. Lacticin 3147 - Biosynthesis, Molecular Analysis, Immunity, Bioengineering and Applications. Current Protein & Peptide Science <i>Antimicrob Agents Chemother</i> volume 13, pages 193-204.</td>
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    </tr>
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      <tr>
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      <td class="references_left">[6]</td>
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      <td class="references_right">Dougherty et al., 1998. Sequence and analysis of the 60 kb conjugative, bacteriocin-producing plasmid pMRC01 from Lactococcus lactis DPC3147 <i>Mol. Microbiol.</i> 29 (4), 10291038</td>
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    </tr>
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            <tr>
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      <td class="references_left">[7]</td>
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      <td class="references_right">Silkin, L. et al., 2008. Spermicidal bacteriocins: Lacticin 3147 and subtilosin A <i>Bioorganic & Medicinal Chemistry Letters</i> 18  3103–3106 Spermicidal</td>
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    </tr>
 +
      <tr>
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      <td class="references_left">[8]</td>
 +
      <td class="references_right">Xiaowen Liu, et al,. 2006.  Engineered vaginal lactobacillus strain for mucosal delivery of the human immunodeficiency virus inhibitor cyanovirin-N. <i>Antimicrobial agents and chemotherapy</i> 50(10), 3250-3259.</td>
 +
    </tr>
 +
      <tr>
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      <td class="references_left">[9]</td>
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      <td class="references_right">Bao, Sujin, et al.2013 "Distribution dynamics of recombinant Lactobacillus in the gastrointestinal tract of neonatal rats." PloS one 8.3 (2013): e60007.</td>
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    </tr>
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  </table>
 
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Revision as of 14:05, 14 October 2018

Wetlab - Peptides

Design


General design

Each of our constructions contained RpsU promoter [8] which is a Lactobacillus jensenii strong promoter. This RpsU sequence also contains the putative sequence for the RBS. We added spacers to all of our constructions to unable easier use of the sequence and separation of the different genes of the sequences. We used two Terminators to our sequences :BBa_B0014 & BBa_B0015 to ensure the stopping of the transcription. Our constructions were assembled in the Plem415 vector by Gibson Assembly method. Plem 415 is a plasmid that works in Lactobacilli species but it’s not specific to L. jensenii [9].

Figure 3 : legend of the designs

SubtilosinA


Figure 4 : Design of the sequence coding the Subtilosin protein with the RpsU promoter.


Figure 5 : Design of the sequence coding the Iron Sulfur Cluster with the RpsU promoter.

Lacticin 3147

This circuit was made from 2 native genes of Lactococcus Lactis ltA1 and ltnA that express Lacticin peptide. Also, the design contains Lacticin-post-transcriptional regulator ltM1 and M2. A promoter orthogonal was used : ptsH and differents spacer taken from igem_parts.

Figure 6 : Design of the sequence coding the Lacticin 3147

LL-37

The design of LL-37 is simpler than the one of the other peptides. Indeed, the protein is coded only with the well-named gene LL-37.

Figure 7 : Design of the sequence coding the LL-37 protein with the RpsU promoter.

Figure 8 : Design of the sequence coding the LL-37 protein with the pHyperSpank promoter.

References
[1] Nguyen LT1, Haney EF, Vogel HJ. 2011. The expanding scope of antimicrobial peptide structures and their modes of action. Trends Biotechnol Vol 29, 464–472..
[2] Tanphaichitr, Nongnuj et al. 2018. Potential Use of Antimicrobial Peptides as Vaginal Spermicides/Microbicides. Pharmaceuticals 9.1 (2016): 13.
[3] Cavera, V. L., Volski, A., & Chikindas, M. L. 2015. The natural antimicrobial subtilosin A synergizes with lauramide arginine ethyl ester (LAE), ε-poly-l-lysine (polylysine), clindamycin phosphate and metronidazole, against the vaginal pathogen Gardnerella vaginalis. Probiotics and antimicrobial proteins. 7(2), 164-171.
[4] Paul M. Himes, Scott E. Allen, Sungwon Hwang, and Albert A. Bowers. 2016.Production of Sactipeptides in Escherichia coli: Probing the Substrate Promiscuity of Subtilosin A Biosynthesis. ACS Chemical Biology. 11 (6), 1737-1744.
[5] Srinivas Suda et al., 2012. Lacticin 3147 - Biosynthesis, Molecular Analysis, Immunity, Bioengineering and Applications. Current Protein & Peptide Science Antimicrob Agents Chemother volume 13, pages 193-204.
[6] Dougherty et al., 1998. Sequence and analysis of the 60 kb conjugative, bacteriocin-producing plasmid pMRC01 from Lactococcus lactis DPC3147 Mol. Microbiol. 29 (4), 10291038
[7] Silkin, L. et al., 2008. Spermicidal bacteriocins: Lacticin 3147 and subtilosin A Bioorganic & Medicinal Chemistry Letters 18 3103–3106 Spermicidal
[8] Xiaowen Liu, et al,. 2006. Engineered vaginal lactobacillus strain for mucosal delivery of the human immunodeficiency virus inhibitor cyanovirin-N. Antimicrobial agents and chemotherapy 50(10), 3250-3259.
[9] Bao, Sujin, et al.2013 "Distribution dynamics of recombinant Lactobacillus in the gastrointestinal tract of neonatal rats." PloS one 8.3 (2013): e60007.

Sponsors



Contact us

igem.assomtp@gmail.com

Centre de Biochimie Structurale, 29 Rue de Navacelles, 34090 Montpellier, France