Part:BBa_K2666001

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 (more information). 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]

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.