Difference between revisions of "Team:Montpellier/WetLab Peptides"
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<h1>Wetlab - Peptides</h1> | <h1>Wetlab - Peptides</h1> | ||
| − | <h2>Design</h2> | + | <h2>Design</h2><hr/> |
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<h3>General design</h3> | <h3>General design</h3> | ||
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<p>Each of our constructions contained RpsU promoter <a class="lien" href="#references">[1]</a> which is a <i>Lactobacillus jensenii</i> strong promoter. This RpsU sequence also contains the putative sequence for the RBS. | <p>Each of our constructions contained RpsU promoter <a class="lien" href="#references">[1]</a> which is a <i>Lactobacillus jensenii</i> 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. | 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 | + | Our constructions were assembled in the pLEM415 vector by Gibson Assembly method. pLEM415 is a plasmid that works in <i>Lactobacilli</i> species but it’s not specific to <i>L. jensenii</i> <a class="lien" href="#references">[2]</a>. |
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| − | + | <center><img class="legend" src="https://static.igem.org/mediawiki/2018/7/7f/T--Montpellier--design_legende3_montpellier.png"></center> | |
| − | + | <figcaption> <span class="underline">Figure 1:</span> Caption of the designs.</figcaption> | |
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<h3>LL-37</h3> | <h3>LL-37</h3> | ||
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<center><img class="design" src="https://static.igem.org/mediawiki/2018/c/c4/T--Montpellier--LL37_design2_mtp.png"></center> | <center><img class="design" src="https://static.igem.org/mediawiki/2018/c/c4/T--Montpellier--LL37_design2_mtp.png"></center> | ||
| − | < | + | <figcaption><span class="underline">Figure 2:</span> Design of the sequence coding the LL-37 protein with the RpsU promoter.</figcaption> |
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<center><img class="design" src="https://static.igem.org/mediawiki/2018/4/4a/T--Montpellier--LL37_hyperspank_mtp.png"></center> | <center><img class="design" src="https://static.igem.org/mediawiki/2018/4/4a/T--Montpellier--LL37_hyperspank_mtp.png"></center> | ||
| − | < | + | <figcaption><span class="underline">Figure 3:</span> Design of the sequence coding the LL-37 protein with the pHyperSpank promoter.</figcaption> |
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| + | <h3>SubtilosinA</h3> | ||
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| + | <center><img class="design" src="https://static.igem.org/mediawiki/2018/3/3b/T--Montpellier--sboaalba_montpellier.png"></center><br/> | ||
| + | <figcaption><span class="underline">Figure 4:</span> Design of the sequence coding the Subtilosin protein with the RpsU promoter.</figcaption> | ||
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| + | <center><img class="design" src="https://static.igem.org/mediawiki/2018/c/cc/T--Montpellier--sufsboa_design_net_mtp.png"></center><br/> | ||
| + | <figcaption><span class="underline">Figure 5:</span> Design of the sequence coding the Iron Sulfur Cluster with the RpsU promoter.</figcaption> | ||
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| + | <h3>Lacticin 3147</h3> | ||
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| + | <p>This circuit was made from 2 native genes of <i>Lactococcus lactis</i> 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.</p> | ||
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| + | <center><img class="design" src="https://static.igem.org/mediawiki/2018/1/17/T--Montpellier--lacticin_design_net_mtp.png"></center> | ||
| + | <figcaption><span class="underline">Figure 6:</span> Design of the sequence coding the Lacticin 3147</figcaption> | ||
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</section> | </section> | ||
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<section class="references" id="references"> | <section class="references" id="references"> | ||
<table class="references_table"> | <table class="references_table"> | ||
Revision as of 08:54, 15 October 2018
Wetlab - Peptides
Design
General design
Each of our constructions contained RpsU promoter [1] 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. pLEM415 is a plasmid that works in Lactobacilli species but it’s not specific to L. jensenii [2].
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.
SubtilosinA
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.
| References | |
|---|---|
| [1] | 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. |
| [2] | Bao, Sujin, et al.2013 "Distribution dynamics of recombinant Lactobacillus in the gastrointestinal tract of neonatal rats." PloS one 8.3 (2013): e60007. |