Yeast Leu2 and Ura3 proteins

Jianhua Yu yu19 at purdue.edu
Thu Jan 10 12:58:58 EST 2002


Hello,

I am making mutable LEU2 and URA3 in S. cerevisiae, i.e. I put
transposon in these two genes and then I hope they can recover function
after transposon jumps away. I have tried 5'UTR first, however,
unfortunately, the transposon I chose seems to have yeast promoter in
both orientations. I have also tried several sites in the middle of
genes, but both genes can not recover function after transposon jumps
away. The problem for this is that transposon leaves  "footprints" and
the sites I chose can not tolerate adding or deletion of extra codons
("footprints").

Does anybody know these two genes very well and can tell me which site
in the middle of gene can tolerate extra codons. i.e. adding or deletion
of some codons at these sites won't affect gene function because these
sites are not functional domain? Or does anybody can inform me how to
solve promoter problem at 5'UTR except mutation, for the mutation may
cause transposon inactive?

Any suggestion is appreciated.

Best,

Jianhua Yu






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