We are trying to clone a mammalian cDNA by complementation of a yeast mutation.
We have a rat gut cDNA library in the vector pYES2 in which the cDNAs are under
the control of the GAL1 promoter. The need to have galactose present is a
problem fro selection. We were wondering if anyone has a mammalian cDNA
library with some other promoter, which they would be willing to supply to us.
Ideally, the cDNA would be from some tissue rich in mitochondria, such as liver
or heart, but some other tissue would do. The yeast strain is mutant for ura3
and his4. If anyone can help us we would be most grateful and would gladly
supply our library in return.
I found that when I put the yeast PRO1 gene (without its own promoter) under
the control of the GAL1 promoter in pYES2 it was able to transform a pro1
mutant to proline prototrophy on glucose or galactose media. I wonder if this
means that we don't need to have galactose present to get expression of the
cDNAs of the library in any strain or if it was due to some property of the
pro1 mutant we were using. eg. perhaps the strain was deficient in one of the
components involved in catabolite repression. Does anyone have experience with
the use of the GAL1 promoter?
La Trobe University
E-mail: biosgm at lure.latrobe.edu.au
Fax: 009 613 479 2467