B6. Gall gene encodes enzyme galactokinase which phosphorylates sugar galactose

Question

B6. Gall gene encodes enzyme galactokinase which phosphorylates sugar galactose for catabolism (utilized for energy). Gall gene is activated when sugar glucose is absent in the medium. Gall gene is not activated by galactose when glucose is present. This is because transcription factor MIGI binds to MIGI site in the Gall promoter and recruits TUPI to repress the Gall gene in yeast when glucose is present in the sugar galactose is present and medium. (1) How will you determine that MIGI binds to MIG1 site only when glucose is present in the medium and not present when galactose (no glucose) in the medium. (2) How will you establish that MIG1 transcription factor forms a complex with TUPL?

Explanation / Answer

In order to determine the transcriptional factor attached to the regulatory protein we can use reporter genes. some commonly used reported genes are GFP, GUS and Lusiferase. They will be tagged to the regulatory elements of the gene of our interest to coexpress with it. They may produce visuval avidence for the protein expression. Seperate set of experiments could be conducted one is with in the presence of glucose and galactose and in the other is only galactose in the media to determine the transcriptional factor attachment to the promoter.

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