problems with two-hybrid

ZX xuez at esvax.dnet.dupont.com
Tue Dec 3 10:32:50 EST 1996


>
I have been screening a cDNA library for interacting proteins using 
the
GAL4 system. So far I have found 10 yeast clones containing potentiall
interactor coding plasmids as they all go blue in the lacZ assay and 
also grow
very well on minus histidin plates. Four of them contained no cDNA 
insert,
the other 6 did not go blue nor grow on minus histidin after 
retransformation
into the same yeast strain. Can anybody help me with an explanation? I 
have
some clones either going blue or growing on minus histidin but I never
investigated those further. Is it possible or rather likely that one 
gets
false-positives for both the reportert genes? I would be very grateful 
in case
somebody comes up with similiar experience or advice.

Thanks for now.    inge

When you get the plasmid out, how many E. coli transformants did you
get? and how many you tested?  Sometimes there are multiple types of 
GAD plasmids in a yeast colony, and may be you should double check.  

Zhixiong Xue
DuPont Central Research



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