Difference between revisions of "Part:BBa K2666004"

Revision as of 10:09, 10 October 2018

scFv LaM-4 with signal peptid YncM 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 B. subtilis specific [2]. Figure 1 illustrates the detailed design.

File:T--Montpellier--interlab od600 mtp.PNG

Figure 1 : Construct design. The sequence that encode scFv LaM-4 with a SP YncM 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.

II. Proof of function

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] Brockmeier, Ulf, et al. "Systematic screening of all signal peptides from Bacillus subtilis: a powerful strategy in optimizing heterologous protein secretion in Gram-positive bacteria." Journal of molecular biology 362.3 (2006): 393-402.

[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 prod