Gene knockout in E.coli

P.R. Alefounder mbpra at s-crim1.dl.ac.uk
Wed Aug 14 06:19:19 EST 1996


In article <morby-1408960934540001 at mnemosyne2.bioc.cf.ac.uk>, morby at cf.ac.uk (Andy Morby) writes:
|> Has anyone seen a recent paper on an E.coli strain which is permissive for
|> gene knockout by homologous recombination i.e by introduction of a neo
|> cassette etc.  I'm sure it exists but can't seem to source it by the usual
|> channels.
|> 
|>       Thanks
|>                Andy Morby

I understand this can be done using a recD strain. This allows
transformation with linear DNA containing your gene with the neo
cassette insert, or some other selectable marker. The recD gene 
encodes a subunit of the exonuclease that would otherwise degrade
any linear DNA before homologous recombination had a chance to 
take place. I have not tried this method myself.

Peter Alefounder.



More information about the Methods mailing list