Part:BBa_K2666000

scFv LaM4 with signal peptid CbsA with a L.jensenii specific strong promoter

I. Part BBa K2666000 : Function

The Montpellier iGEM team 2018 designed a construction that produce and secrete LaM-4, a nanobody against RFP and mcherry in Lactobacillus jensenii (more information). For that, we used the promoter RpsU, a L.jensenii specific strong promoter [1]. The signal peptid is L. jensenii specific [2]. Figure 1 illustrates the detailed design.

Figure 1 : Construct design. The sequence that encode scFv LaM-4 with a SP CsbA and produce by a strong promoter RpsU.

LaM4 is used as a control [3]. It is an antibody against RFP & mcherry. This control allow us to check the efficiency of the promoter and the signal peptid.

III. Design Considerations

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] Expression of Human Immunodeficiency Virus Type 1 Neutralizing Antibody Fragments Using Human Vaginal Lactobacillus ; Engineered Vaginal Lactobacillus Strain for Mucosal Delivery of the Human Immunodeficiency Virus Inhibitor Cyanovirin-N [3] Fridy, P. C., Li, Y., Keegan, S., Thompson, M. K., Nudelman, I., Scheid, J. F., ... & Rout, M. P. (2014). A robust pipeline for rapid production of versatile nanobody repertoires. Nature methods, 11(12), 1253.