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> ******************************************************************
