yeast transformations

Tue Sep 27 14:26:18 EST 1994

                   Ohio University Electronic Communication

   Date:  27-Sep-1994 03:01pm EST

     To:  Remote Addressee                     ( _MX%"yeast at" )

   From:  Tejal Bhatt                                Dept:  Biological Sciences
          BHATT                                    Tel No:  (614) 593-4635

Subject:  yeast transformations

Hi. I have been attempting to transform a strain of S. cervisiae, known as 
ZM137, with a construct I made to express a certain protein. The strain has a 
mutation in the triose phosphate isomerase and is unable to metabolize glucose, 
so it is grown on media which contains galactose, glycerol, and ethanol. The 
vector contains the triose phosphate isomerase gene; allowing transformants to 
be selected on glucose containg media. Besides being a poor growing strain, it 
is not transformable using standard lithium acetate methods (believe me I have 
tried!). I have also tried electroporation and spheroplasting, both without      
success. I have tried every protocol I can find- from Methods in Enzymology to
Current protocols in Molecular biology. I have bought a spheroplasting kit, 
which is not working either. If anyone has any suggestions or protocols which 
you doubt I have tried; I would appreciate your input. 
      Tejal Bhatt

Received:  27-Sep-1994 03:28pm

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