selecting against petite
James J. Campanella
jjc4 at po.CWRU.Edu
Wed May 5 16:03:01 EST 1993
I hope that this is not one of the boards that's poorly used. Here
is my dilemma. I have transformed a gene that I am interested in
into a petite strain of yeast (1893... I used the strain for
selection purposes since it is auxotrophic for both Phe and Ura, and
I needed both those markers). The problem is that the strain grows
very slowly-- this is probably due to a combination of the fact that
the line is petite and the gene I transformed in is not friendly to
yeast physiology. It takes about a week to get the transformed strain
up to an OD of 1.5-- and that turns out to be stationary phase, since
I have yet to record greater growth. At any rate, that speed of growth
is simply not practical for experiments.
Does anyone have any suggestions on how to grow up this transformant
and maybe select against petite so that I can get rid of at least one
factor that's probably slowing growth.
Does anyone have any suggestions at all on speeding up growth?
I've been growing the transformants on MvDex supplemented with amino
acids and selecting for my plasmid of interest by just leaving out
URA. I've tried growing on YPD, but without selection I lose my
plasmid and my gene along with it.
Any help would be appreciated. Thanks.
James Campanella : "I started out with nothing, and now,
Case Western U. : I have worked my way up to a state
Biology Dept. : of abject poverty."
Cleveland, OH : - Monkey Business, Groucho Marx
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