We are also having this problem but with the one-hybrid system. Yeast
(YWAM2) harboring our CEN reporter plasmid are not his+, at least when
streaked out on his- media. When we transform with our library, however,
we get thousands of his+ colonies. Replica plating the transformants to
150mM 3AT retarded the growth of all of the colonies, but did not reduce
the number of his+ colonies. Being rather new to this, we are not sure
how to get around this problem. Would changing strains help? How does
3AT inhibit background his expression anyway?
Any advice you could give would be appreciated. Sorry we can't help you out!!
Please reply to:
kucharck at mail.med.upenn.edu
