Difference between revisions of "Part:BBa K2666001"
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The Montpellier iGEM team 2018 designed a construction that produce LL-37, an antimicrobial peptide from human macrophages and leukocytes in Lactobacillus jensenii (http://2018.igem.org/Team:Montpellier/Peptides) | The Montpellier iGEM team 2018 designed a construction that produce LL-37, an antimicrobial peptide from human macrophages and leukocytes in Lactobacillus jensenii (http://2018.igem.org/Team:Montpellier/Peptides) | ||
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For that, we used the promoter RpsU, a L.jensenii specific strong promoter [1]. This RpsU sequence also contains the putative sequence for the RBS. Figure 1 illustrates the detailed design. | For that, we used the promoter RpsU, a L.jensenii specific strong promoter [1]. This RpsU sequence also contains the putative sequence for the RBS. Figure 1 illustrates the detailed design. | ||
| + | [[File:T--Montpellier--LL-37-rpsU design mtp.PNG|550px]] | ||
| − | Figure 1 : Construct design. The sequence that encode LL-37 with a strong promoter RpsU. | + | '''Figure 1''' : Construct design. The sequence that encode LL-37 with a strong promoter RpsU. |
LL-37 has shown effect in vivo in mice. Females naturally cycling to estrous phase failed to become pregnant when in injected with LL-37 and sperm. LL-37 also showed in vitro effect on human sperm which makes it a very good potential peptide for contraception. [2] | LL-37 has shown effect in vivo in mice. Females naturally cycling to estrous phase failed to become pregnant when in injected with LL-37 and sperm. LL-37 also showed in vitro effect on human sperm which makes it a very good potential peptide for contraception. [2] | ||
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We used a strong promoter to produce a lot of LL-37. Lactobacillus is not supposed to be sensitive to LL-37 (reference). | We used a strong promoter to produce a lot of LL-37. Lactobacillus is not supposed to be sensitive to LL-37 (reference). | ||
| − | 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. |
== Reference: == | == Reference: == | ||
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
| − | <partinfo> | + | <partinfo>BBa_K2666001 SequenceAndFeatures</partinfo> |
Latest revision as of 12:38, 17 October 2018
Contents
LL-37 with L.jensenii specific strong promoter RpsU
I. Part BBa K2666001 : Function
The Montpellier iGEM team 2018 designed a construction that produce LL-37, an antimicrobial peptide from human macrophages and leukocytes in Lactobacillus jensenii (http://2018.igem.org/Team:Montpellier/Peptides)
For that, we used the promoter RpsU, a L.jensenii specific strong promoter [1]. This RpsU sequence also contains the putative sequence for the RBS. Figure 1 illustrates the detailed design.
Figure 1 : Construct design. The sequence that encode LL-37 with a strong promoter RpsU.
LL-37 has shown effect in vivo in mice. Females naturally cycling to estrous phase failed to become pregnant when in injected with LL-37 and sperm. LL-37 also showed in vitro effect on human sperm which makes it a very good potential peptide for contraception. [2]
II. Proof of function
III. Design Considerations
We used a strong promoter to produce a lot of LL-37. Lactobacillus is not supposed to be sensitive to LL-37 (reference).
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.
Reference:
[1] Bao, Sujin, et al. "Distribution dynamics of recombinant Lactobacillus in the gastrointestinal tract of neonatal rats." PloS one 8.3 (2013): e60007.
[2] Tanphaichitr, Nongnuj et al. 2018. Potential Use of Antimicrobial Peptides as Vaginal Spermicides/Microbicides. Pharmaceuticals 9.1 (2016): 13.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 62
Illegal BamHI site found at 194 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]