IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/08/05
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Testing AHL degradation by aiiA
We are testing B. subtilis strain 168’s ability to degrade AHL by exposing synthetic AHL to the strain and then testing its ability to be recognized by the receiver.
Bacillus subtilis - protease deficient (NPR and APR) derived from strain 168 - grown up in an over night culture of LB.
Synthetic AHL – Sigma K3007 – AHL powder was dissolved in EA buffer and diluted to 10uM concentration
E. coli with T9002 AHL receiver grown up in overnight culture of LB
All plates contain soft agar with 200uL of E. coli AHL receiver mixed-in. Plates 1,2 and 4 contain Bacillus in fresh LB medium, while plate 5 contains Bacillus in it’s overnight LB. Plate 5 was used to test if an excreted product of Bacillus is responsible for the degradation of AHL. The product of the aiiA gene in Bacillus subtilis is most likely not exported.
plates visualized 3 hours after AHL/Bacillus inoculation.....