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			
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Rafael at corona.med.utah.edu		"No por mucho madrugar, amanece
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