High-effeciency mating protocols

Matthew Spottswood spottsy at monod.Biol.McGill.CA
Mon Apr 11 20:31:44 EST 1994


labfw at rascal.med.harvard.edu (Fred Winston) writes:

>Hello:
>	I am trying to do a cross with a mutant strain that is apparently 
>sterile.  I have tried the filter mating protocol published by Hartwell, and 
>that didn't work.  Does anyone know of any other effective methods of mating 
>reluctant strains?  Thanks.
>						Dave Rose 

Hi Dave,
	Let's say that the strain you're trying to mate is sterile, you're
faced with only two possible solutions.  The most obvious is to rescue the
mutation which results in the sterile phenotype with a yeast genomic clone
in an easily "curable" plasmid (i.e. one constructed in a URA3 or LYS2
vector).  Obviously this should restore wild type mating to your strain. 
Of course this assumes that your strain has a single mutation that is
responsible for all the ills which afflict your strain.
	If this doesn't appear feasible then you could try a spheroplast 
fusion (van Solingen and van der Plaat (1977) J. Bac 130(2) pp.946-947) 
which doesn't require the mating reaction to occur. Hopefully this 
information will help.

						Good Luck

						Matt Spottswood  



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