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Vector stability

S. L. Rennie rennie at bldghsc.lan1.umanitoba.ca
Thu Oct 12 13:34:59 EST 1995


In article <45gm0l$40u at news.bu.edu>, harkness at bio.bu.edu (Daniel Harkness)
wrote:

> Hello All,
> 
> Does anyone have any information regarding the maximum number of plasmids
> one can transform into S. cerevisiae?  I have a trp-, his-, ura-, leu-
> strain and would like to transform 4 constructs that carry the 4 different
> markers and of course select on u-, t-, l-, h- medium.  Is there anything
> I should worry about?  The vectors are 2 expression (CEN and 2um), 1 cDNA 
> library (2um), 1 reporter (CEN).
> 
> Thanks,
> Dan Harkness
> Boston University
> Dept. of Cell and Molecular Biology
> 5 Cummington St.
> Boston, MA 02215
> email: harkness at bio.bu.edu
---------------------------------------

I this a 2HS screen?  Maybe you should consider using a system where your
reporter is integrated into the strain and then at least you're down to
three?

We've certainly done three before but this is an in vivo system here and
I'm beginning to wonder about the validity of the results when we start
asking to much of it.

More optimisitcally I'd transform one or two at a time to confirm they're
in before transforming the library.  Try to optimize lib trafo so you
don't get multiples in and try to balance expression of 'test vector' for
optimal interaction.

That's about all I can think of for now, hope this is what you were
looking for and not just useless babble

Good luck 
Sharon

-- 
we're not happy if our yeast ain't blue!




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