K.lactis problems??

Ron Winkler winkler at rulsfb.leidenuniv.nl
Tue Aug 27 17:33:43 EST 1996


Email (YourName) wrote:

>Hi there, has anybody ever had any problems with creating mutants via
>gene replacement in K.lactis?? I've done a couple of experiments by
>electroporating my replacement construct into the cells, and so far only
>approx 0.5% of transformed cells have the expected gene replacement. Is 
>this construct/locus/strain dependant and does anybody have any solutions/
>answers? Thanks.

I have made 4 gene disruptions in K.lactis. 2 were easy, 2 were more
difficult. From what I gather there seems to be a locus specific
effect, but there are also other things that have an effect. But I
don't know what they are. Frequencies are generally low (similar to
yours), but sometimes for some reason or other frequencies are much
higher.

Ron




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