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
******************************************************************
