For a gal4∆ cell that expresses both the bait and prey plasmids, which cells will survive...

For a gal4∆ cell that expresses both the bait and prey plasmids, which cells will survive on galactose containing media?

Expert Solution

Answer: The Y2H (yeast two hybrid) system is used to detect the protein-protein interaction. There is an interaction between two proteins known as bait and prey which leads to activation of the reporter gene.

During the molecular analysis of eukaryotic transcription factor, it was observed that in yeast, Gal4 binds to upstream activation domain and initiates transcription of structural gene in presence of galactose. If this Gal4 is separated into two fragments as N terminal fragment (DNA binding domain) and C terminal fragment (activation domain), if there is only N terminal fragment which binds to the DNA it is incapable of initiating transcription. Hence interactions of both the fragments is necessary for activation of transcription.

There are two plasmid vectors made one with each of the genes i.e. one with DNA binding domain called as bait and another with activation domain called as prey. The host cells are then transformed with the two plasmids, the host cells transformed with both the plasmid together will be able to express the reporter gene, which can be detected on the galactose containing plate.

If the cells are having only one of the plasmids either bait or prey, they would not survive on galactose containing plate.

Whereas cells containing both the plasmids will be able to survive on galactose containing plate.

gladiator answered 2 years ago

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