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 
advance.

Kevin Piers
Dept. Microbiology
University of Washington
Seattle WA 98195

 



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