Homologous Recombination in Yeast

Dan Baggott dbaggott at reed.edu
Thu Jan 25 14:57:36 EST 1996


"Miguel A. RODRIGUEZ-GABRIEL" <MARODRIGUEZ at mvax.cbm.uam.es> wrote:
>Dear Yeast Netters,
>
>    I have one question (maybe more!) concerning homologous recombination in
>Saccharomyces cerevisiae. My main questions are:
>
>    1. Minimum homologous fragment size needed for homologous recombination.
>    2. Percentage of homology needed for this homologous recombination.

<snip>

Hi Miguel,

I'm not sure if this is what you had in mind but check out:  Schiestl and
Petes. 1991. Integration of DNA fragments by illegitimate recombination in
Saccharomyces cerevisiae. PNAS 88:7585-7589.

Among other things, they conclude that "four base pairs of homology are
sufficient for some types of mitotic recombination."  One of those papers that
blows my mind as to what you can do in molec. biology and get away with.  They
transformed DNA fragments generated by BHI digestion in the presence of the BHI
enzyme and get selective integration into genomic BHI sites.  Without the
enzyme, they integrated only into GATC sites.  Pretty amazing...  Maybe we
should be thinking a little harder when we transform yeast with digests without
inactivating the restriction enzyme.

Dan Baggott
Reed College Biology Dept.
Portland, OR





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