The lab in which you work wants to use a yeast two-hybrid (Y2H) assay to identif

Question

The lab in which you work wants to use a yeast two-hybrid (Y2H) assay to identify binding partners (prey) for a newly discovered yeast protein (bait). Your lab's Y2H system uses the Gal4 transcription factor. You begin by testing the Y2H system with a pair of proteins that exhibit a strong, direct binding interaction in several in vitro binding assays. Unfortunately, you do not see any surviving colonies. You know that the commercial Y2H system yoú used in your experiment is reliable and has been shown to work with bait and prey positive controls. Choose the statements that are reasonable explanations for the failure of your Y2H experiment. Fusing the bait or prey to portions of Gal4 resulted in loss of bait and prey binding due to steric hindrance. The Y2H system lacked necessary cofactors or allosteric effectors required for bait and prey binding. The Gal4 activation domain failed to bind to RNA polymerase. One or both of the fusion proteins did not localize to the nucleus. The expression levels of the fused bait or prey proteins were too low.

Explanation / Answer

Answer) multiples choices can be:

-Fusing the bait or prey to portions of Gal4 resulted in loss of bait and prey binding due to steric hinderance.

-The Gal4 activayion failed to bind RNA polymerase.

- The expression level of fused bait and prei is low.

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