Seeking advice from yeast experts

Warren Lushia walush0 at ukcc.uky.edu
Mon Sep 30 09:44:25 EST 1996


hello yeast-netters,
I am currently looking into the possibility of developing a yeast 
packaging system to package the virus I am working with (tobacco 
mosaic virus).....we are looking at disassembly of the virus, and are 
have made mutants to examine our hypothesis.  However, since these 
mutants are non-replicating, they are not infectious, and we need to 
package them another way.  This virus needs only its coat protein and 
its RNA and it will self assemble....however packaging the virus in 
vitro has many problems, not the least of which is producing in vitro 
a sufficient amount of transcript for the procedure to work.  However, 
a collegue who works with both TMV and has some experince with yeast 
seems to think a yeast system might do the job....the viral coat 
protein would be produced by one vector, and transcripts to be 
packaged would be produced by another (ideally under the control of 2 
different inducible promoters, so we could optimize the system).  The 
virus does not need to replicate, all that would be needed is coat 
protein and genomic transcripts, and the 2 should form particles.  
Since I have 0 experience with yeast systems, I am looking for advice 
on how to begin examining whether this will work, and what plasmids or 
strains you would recommend.  I have a vector with the Gal1 promoter 
and an offer for another with an alcohol oxidase promoter from 
Biosource, which I would try to use in saccharomyces.  I'm not sure if 
this is the best way to go, or if a pichia system may work better.  
Any help comments advice warnings would be appreciated....thanks!  
warren..



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