Team:EPF Lausanne/Calendar/21 September 2013

From 2013.igem.org

Taxi.Coli: Smart Drug Delivery iGEM EPFL

Header

Cell Surface Display
New Strategy
- We had to repeat the PCR and Gibson reactions for the new strategy INP_streptavidin_YFP and INP_YFP_streptavidin since the PCR and Gibson performed the week before didn't work.
Western Blot
- We repeated the WB experiment with a new rabbit anti-streptavidin antibody conjugated to HRP. This time put on the gel the cell pellets but also the cell supernatants.
- The gel was not of good quality so the results were not good at all.

Sensing-Effector

Measuring GFP expression of the hya and the cad promoters
-We incubated the cells in different media (acidic, neutral and alkaline) in order to find out when the promoters are induced. The results were measured with a plate reader. They showed that the bacteria at the low pH died very quickly and didn’t express much GFP. The ones at neutral pH didn’t express much GFP either. Only the bacteria in the basic medium with Have some sort of positive induction. This seemed strange since both promoters should induce expression at lower pH and not at basic pH.

Western blot of the cell lysates/supernatants (bacteria transformed with MMP2 and GelE).