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pombe GFP vectors?

S L Forsburg forsburg at nospamsalk.edu
Sun May 17 08:02:17 EST 1998


> From: Nick <nick_owen at wo*ls.demon.co.uk>
> 
> Subject: pGP110 Vector - anyone?
> 
> Dear All,
> 
> In the following reference:
> 
> Methods Enzymol 1997;283:459-471
> 
> Use of green fluorescent protein for intracellular protein
> localization in living fission yeast cells.
> Nabeshima K, Saitoh S, Yanagida M
> 
> they refer to a vector of expression in S.pombe that has the LEU2
> marker and nmt promoter and stops. The cloning site has SalI, BamHI
> and NotI restriction sites which is absolutley perfect for the linker
> primers I have got for a number of cDNA's that I have amplified. By
> using these sites it is possible to use the vector to 3' tag the cDNA
> with the GFP ORF.
> I was wondering if anyone out there has this pGP110 vector for
> expression in yeast?


I don't know about the vector you mention.  However my lab has 
recently constructed general purpose S. pombe GFP vectors in
the REP4X background, with  compatible polylinkers to the
GST and HA versions we published last year. Our vectors 
offer N-terminal or C-terminal fusions, ura4+, and the full strength
nmt1 and terminator.  We are slowly building a family of such
tagging vectors all of which use the same polylinker, thus
allowing a cDNA to easily swapped between different tags.  We
have submitted our plasmids to the ATCC for distribution. 

More information, including ATCC accession numbers, 
 is available on our pombe vector database 
http://flosun.salk.edu/~forsburg/vectors.html


-- 
-susan
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S L Forsburg, PhD  forsburg at salk.edu
Molecular Biology and Virology Lab          
The Salk Institute, La Jolla CA 
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