KAR1 transfer of YACs from AB1380
tomlinson at pplros.demon.co.uk
Thu Jan 18 05:18:29 EST 1996
I've been trying to use the protocol of Spencer et al (Genomics 22:118,
to transfer a 400Kbp YAC from AB1380 to YPH925 (Kar1).Basically the
advantage of the phenomenon of cytoduction and the rare "chromoductant"
I have two questions/observations that, hopefully, someone can help me
1. The YPH925 stab that arrived from ATCC was clearly non-clonal as it
and red (ade-) areas. On streaking it was clear that I got white, red and
colonies. I have assumed that this is some effect of the Kar1 mutation, is
it? Is it possible to get a stable clonal culture of such a mutation? If I
colony purify, is there a simple way of testing for Kar1?
2. Upon mating my YAC strain (called HP8 for ease of writing/typing) and
on ura drop out plates plus cycloheximide, I get a very high spontaneous
rate of Cy resistance. I do not see this with another strain containing
same YAC and made by traditional mating (to YPH499 or 500, I can never
mating types!). Is this something that others have seen? I don't think
in the paper. If AB1380 gives a high CyR background it makes the
hell of a lot more hassle.
Thanks in advance
PPL therapeutics Ltd
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