Dear yeast netters:
I am trying to complement a yeast mutant with arabidopsis cDNA clones. The
arabidopsis cDNA library is constructed in lambda YES shuttle vector. YES has
lac and gal promoters that drive expression in E. Coli and yeast respectively.
Lac and gal promoters are flanking the cloned cDNA inserts.on the opposite
ends.Here comes the puzzling part: one of the arabidopsis clones that
complemented the yeast mutant has the cDNA insert in the orientation that it
is being driven by the lac promoter ie beginning of the cDNA is next to lac
promoter. How is this cDNA able to complement the yeast mutant. Does lac
promoter has any activity in yeast? or alternativley antisense mRNA is being
produced due to gal promoter activity. What is the most plausible explanation ?
Any replies and suggestions will be highly appreciated.
Yogesh Sharma
