E.coli "gene knockout": help
Rafael Maldonado
rafael at corona
Sun Mar 12 00:40:44 EST 1995
On 10 Mar 1995, Richard_Heath wrote:
>
> Another way to knock the gene out is to put an antibiotic resistance marker
> into your cloned gene in a plasmid, then transform with linear DNA obtained
> from this new plasmid. Homologous recombination should insert the
> gene::antibiotic marker construct into the chromosome in place of the original
> gene. This method has been used successfuly in our lab recently - don't have
> any refs to hand right now though.
But you need use a rec(something)- strain. Others are not capable of
linear DNA transfromation. I don't remember the rec gene, may be recB
scb? Can someone pint us to this system?
Rafa
__________________________________________________________________________
Rafael Maldonado "You gave to a boy the job
Department of Human Genetics of a man. He's dead."
University of Utah
Salt Lake City, Utah 84112. USA. John Wayne
Rafael at genetics.med.utah.edu
Rafael at corona.med.utah.edu "No por mucho madrugar, amanece
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