problems with two-hybrid

weber at
Tue Dec 3 05:30:45 EST 1996

hi everybody!

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  

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