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vacuum infiltration of yeast

Kevin Piers klpiers at U.WASHINGTON.EDU
Tue Oct 25 13:20:41 EST 1994

Hello oh fellow yeasters,

I have a question that may initially sound bizarre, but I am relatively 
new in this field.  I am trying to establish a system by which I can 
routinely and consistently transform yeast using agrobacterium as a donor 
organism.  I have all the appropriate vectors set up and have been able 
to obtain transformants in this way.  Due to the fact that S. cerevisiae 
is relatively "non-sticky" and that cell to cell contact is necessary for 
DNA transfer in this system, I have promoted such interactions by vaccuum 
infiltrating these organisms together.  That is, once they are mixed and 
place on a filter, I subject them to a time in a vacuum. This treatment 
results in a higher transformation frequency.  Now, FINALLY, the 
question:  Has anyone ever subjected yeast to vacuum infiltration and if 
so, under what circumstances and what effects did it have on the cell.  
Although I am running the proper controls, I am slightly concerned about 
the effects of such a treatment on the yeast cells.  Any information 
would be greatly appreciated and can be sent directly to me.  Thanks in 

Kevin Piers
Dept. Microbiology
University of Washington
Seattle WA 98195


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