[Microbiology] Re: COUNTERSELECTABLE MARKERS

Tom Knight tk at mit.edu
Sun Feb 12 18:39:28 EST 2006


Mingfei Shao <mfshao at gmail.com> writes:
> Any one can recommend me some COUNTERSELECTABLE MARKERS for a gram positive
> bacteria.
> I know SacB is a common one for gram negtive bacteria and can lead to many
> useful application.
> However I am now studying a strain from Bacillus spp. is there any
> counterpart  of SacB in gram positive bacterium?

You might want to check out the PheS mutant described by Peter Kast:

Kast P
pKSS--a second-generation general purpose cloning vector for efficient
positive selection of recombinant clones.
Gene. 1994 Jan 28;138(1-2):109-14  PMID: 8125286

You probably would need to modify the gram+ version of pheS.
I found it was important to control the pH of the medium containing
p-chloro phenylalanine.




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