Paris/Results
From 2007.igem.org
Contents |
Experimental results
Survival of dapA deletant
If we want our system to be robust, the germ line cells should be able to live as long as possible when deprived of DAP. They may indeed face such a feat within our system if their is not enough somatic cells at a given time.
Our dapA deletant strain was grown over night in LB + 300µM DAP. A culture was then launched diluting 100 time the ON culture in LB (+ Kanamycin). 100µL were plated each hour and the number of clone determined.
dapA deletant growth
We expect the somatic cells to excrete only a limited amount of DAP, it would thus be nice if our auxotroph strain could grow on limited amount of metabolite. Over night culture were diluted 100 time in LB suplemented with different amount of DAP.
DAP excretion by prototroph cells
We want to know if prototroph cells are excreting DAP and in what amount. Instead of doing proper chemical dosages which are expensive, we had the idea to make a biological measurments.
Prototroph MG1655 strain was grown in LB, and the medium was recovered at different stages of the culture and purified. The dapA deletant strain was then grown in the recovered medium. The idea is that comparing the growth curve of this experiment with growth curves of dapA deletant in differently supplemented LB medium, should alow us to determine approimatively the amount of DAP present in the recovered medium.
