is there anyone have experience with the bacillus subtilis? I have got a plasmid that can multify in E.coli both with Amp or Chloramphenicol resistance. the plasmid grow well when Amp is added, but is not good when Chloramphenicol is added 5ug/ml, the transformed colony will not bee seen untill 2 days grow in 37C. is there anything wrong?

in antibiotic medium, bacteria always grow slowly inspite of resistance gene contain
5 g/l is a low concentration
i think y should spread on non-antibiotic medium first
if it still no result ,y much check your plasmid by pcr
hope y will success

hi,

B.subtilis should be Amp resistant because it has a lactamase.

We also use 5µg Cm/ml for selection of transformed B.subtilis and this works.

Sometimes colonies of transformed B.subtilis occur 1,5-2 days of incubation.

To be shure you should reisolate the plasmid from Bacillus (incubate with Lysozyme before using a plasmid isolating kit)

good luck