Homologous Recombination in Yeast

Soon A. Lee s.lee at sbsu1.auckland.ac.nz
Mon Jan 29 23:59:11 EST 1996

In article <Roof-2601961651270001 at roof.med.upenn.edu>,
Roof at mail.med.upenn.edu (David Roof) wrote:

> Just 45 bp at each end of a selectable marker is enough to direct
> homologous recombination, provided the original locus for the selectable
> marker has been deleted.  See Baudin et al, NAR 21:3329-3330, 1993.  This
> allows direct gene deletion using a PCR fragment.
> David Roof
The question I think is, how much homology is needed if the marker gene is not deleted.  I have been having trouble (ie mainly int. occurs at the marker locus) with constructs made with a total of 1 kb flanking homology - does anyone know of a comparitive study done on this question?  Also are shorter markers better (more specific) than longer markers, eg TRP1 vs LEU2?
Colin MacDiarmid
macdiarm at sbsu1.auckland.ac.nz

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