chacko@hpuxa.ircc.ohio-state.edu (George W. Chacko) (08/05/90)
If anyone has had experience with Tn5 mutagenesis in Alcaligenes sp. (I'm especially interested in Alcaligenes fecalis) I'd greatly appreciate their comments on this problem. I'm using pGS9 as the source of Tn5 (pGS9 encodes mob and tra functions and also chloramphenicol resistance) mating with Alcaligenes fecalis in a drug free enriched medium (LB) and selecting for kan resistance encoded by Tn5 on a minimal medium for ALcaligenes. The strain of A.fecalis I'm using is kan sensitive. If anyone has a protocol that's worked for them I'd like to get a copy. I'll summarise replies and post if I get enough responses. Thanks George