13. Gene Regulation in Eukaryotes

Which of the following is the purpose of the GAL gene system?

The GAL gene system is activated in which of the following conditions?

When the GAL gene system is activated, galactose binds to the UAS regulatory region.

Compare and contrast the transcriptional regulation of GAL genes in yeast with that of the lac genes in bacteria.

A modification of the two-hybrid system, called the one-hybrid system, is used for identifying proteins that can bind specific DNA sequences. In this method, the DNA sequence to be tested, the bait, is fused to a TATA box to drive expression of a reporter gene. The reporter gene is often chosen to complement a mutant phenotype; for example, a HIS gene may be used in a his⁻ mutant yeast strain. A cDNA library is constructed with the cDNA sequences translationally fused to the GAL4 activation domain and transformed into this yeast strain. Diagram how trans-acting proteins that bind to cis-acting regulatory sequences can be identified using a one-hybrid screen.

Explain how the following mutations would affect transcription of the yeast GAL1 gene in the presence of galactose.

A deletion of one of the four UAS_G elements upstream from the GAL1 gene.

Explain how the following mutations would affect transcription of the yeast GAL1 gene in the presence of galactose.

A mutation within the GAL80 gene that blocks the ability of Gal80 protein to interact with Gal3p.