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I know B does activate A transcriptionally in vivo and AB do form heterdimer both in vivo and in vitro. My concern is that if A has an B-dependent enhancer within its coding sequence and it works with the basal promoter (hsp70 promoter, for example) in the vector, it might be difficult to interpret my data. I just want to make sure I can conclude B somehow stabilizes A at protein level (not activates A transcriptionally) as long as I see the half-life of A is extended.

Thank you very much for you reply. One last quick question, what if my B could also activate A transcriptionally. (I guess I could definitely test that by a RT-PCR or western.) But if that is the case, I would see more A protein in A+B than that in A alone right after the "pulse" phase. Can I get around that issue by looking at the turn-over rate/ half-life of A? Thank you.

Hi CharonY, Thanks for your reply. That is my concern too. If protein complexes break up in a denaturing gel, it should be fine since I will only see a single band that indicates A. However, if I see a high-MW band that indicates AB complex, it's gonna affect my quantification of A's decay since some A are existing in the form of AB complex. So my question is there a way to break up complex on purpose right before running the gel? In that case, I should be able to tell between B stablizing A and B binding A. Thank you

Dear all, I just found this great place to discuss science! I was wondering if anyone has an idea that roughly how many Drosophila genes have a phenotype when mutated? I think I have read it somewhere that only 1/3 of the genes would give a phenotype but am not entirely sure about that. Could anyone please help me out? Thank you