What properties demonstrate that the lac repressor is a protein? Describe the evidence that it indeed serves as a repressor within the operon system.

The locations of numerous lacI⁻ and lacIˢ mutations have been determined within the DNA sequence of the lacI gene. Among these, lacI⁻ mutations were found to occur in the 5′-upstream region of the gene, while lacIˢ mutations were found to occur farther downstream in the gene. Are the locations of the two types of mutations within the gene consistent with what is known about the function of the repressor that is the product of the lacI gene?

The CAP binding site in the lac promoter is the location of positive regulation of gene expression for the operon. Identify what binds at this site to produce positive regulation, under what circumstances binding occurs, and how binding exerts a positive effect.

Describe the experimental rationale that allowed the lac repressor to be isolated.

What role does cAMP play in transcription of lac operon genes? What role does CAP play in transcription of lac operon genes?

How would a cap⁻ mutation that produces an inactive CAP protein affect transcriptional control of the lac operon?

Predict the effect on the inducibility of the lac operon of a mutation that disrupts the function of:

(a) The CRP gene, which encodes the CAP protein

(b) The CAP-binding site within the promoter.

Erythritol, a natural sugar abundant in fruits and fermenting foods, is about 65 percent as sweet as table sugar and has about 95 percent fewer calories. It is 'tooth friendly' and generally devoid of negative side effects as a human consumable product. Pathogenic Brucella strains that catabolize erythritol contain four closely spaced genes, all involved in erythritol metabolism. One of the four genes (eryD) encodes a product that represses the expression of the other three genes. Erythritol catabolism is stimulated by erythritol. Present a simple regulatory model to account for the regulation of erythritol catabolism in Brucella. Does this system appear to be under inducible or repressible control?

Consider the transcription of genes of the lac operon under two conditions: (1) when both glucose and lactose are present and (2) when glucose is absent and lactose is present. Describe the comparative levels of transcription of lac operon genes under these conditions, and explain the molecular basis for the difference.