Suggestions: Vector to knockout E. coli gene.

Richard Koepsel rrk1+ at pitt.edu
Wed Jul 8 09:11:04 EST 1998


Doug,
We've used the phagmid pBIP from Russ Maurer (J Bact. 175; 4260-4262,
1993) with some success.  Dr Maurer is quite helpful and the vector is
supplied with all the revelant strains etc.  We've tried a couple of
other systems and this one seems best.
Rick Koepsel

Doug Stemke wrote:

> I'm looking for recommendations for an E. coli vector for knocking out
> an
> E.coli gene (I have the entire gene already sequenced, I just need to
> make a null mutant). I believe I am looking for something that I can
> confirm knockouts by altering temperature of the culture and
> arresting plasmid replication and then screen the mutants using
> selective
> antibiotic sensitivities. It would also be nice if it had a nice
> polyclonal region as well, but not entirely necessary as I can PCR in
> restriction sites.
>
> Any suggestions?  I've gone through a couple of papers on the topic
> and so
> far haven't found anything I can track down.
>
> Thanks in advance.
>
> Doug Stemke
>
> ******************************************************************
> Douglas J. Stemke, Ph.D.
> Biological Process Technology Institute
> University of Minnesota
> St. Paul, MN
> dstemke at biosci.cbs.umn.edu
> ******************************************************************






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