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Please Help me!! Cloning in Sacchromyces

Arle Kruckeberg arle at biovax.leeds.ac.uk
Fri Mar 8 21:01:38 EST 1996

In article <jpolta-2802961051320001 at medmicro24.med.ohio-state.edu>
jpolta at magnus.acs.ohio-state.edu (Julie Polta) writes:

> I am trying to clone a 2 kb C. albicans gene that just will not clone in
> e. coli (Sure2 or HB101)!!  We have it in two genomic fragments but we can
> not clone the whole genomic fragment or the PCR product of the whole
> gene.  We know that the gene has codon usage typical of c. albicans which
> is to say diff from e. coli.  I have to clone the gene because I want to
> disrupt it with a hisg/URA3 disruption cassette.  
> My question is this:  Can I use sacchromyces to clone the gene and all 
> subsequent manipulations.  I am getting desperate.
> Thanks for your help.
> Chris Ferrer
> cferrer at magnus.acs.ohio-state.edu
Hi Chris--

I can think of 2 examples of constructs that people couldn't make in E.
coli so they transformed the ligation straight into yeast. One was a
construct involving Mal61 (a trpE fusion?) in Corinne Michel's lab and
the other I've forgotten, but also a membrane protein. In any case, it
can work...try it!
Good luck...

a.k.a. Dr. Arthur L. Kruckeberg
 Department of Biochemistry and Molecular Biology
University of Leeds        phone  +44 +113 2333172
Leeds LS2 9JT              FAX    +44 +113 2333167
Great Britain              arle at biovax.leeds.ac.uk

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