How can gene duplication aid in phenotypic divergence? Briefly describe how the lac repressor and catabolite activator protein (CAP) work to regulate expression of the lac operon.
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- 5. Consider the following examples that include both haploids and partial diploids and explain in each scenario a. whether the repressor can bind and regulate expression and b. whether or not the lac operon is expressed. For partial diploids the plasmid is indicated by the F’. I + O +Z + Y + / F’ I+ O + Z + Y + I - O + Z + Y + / F’ I+ O + Z + Y + I - O + Z + Y + I + O c Z + Y +1. Describe which enzymes are required for lactose and tryptophan metabolism in bacteria when lactose and tryptophan, respectively, are (a) present and (b) absent. 2. Contrast positive versus negative regulation of gene expression. Describe the role of the repressor in an inducible system and in a repressible system.1. How does induction control enzyme synthesis using lactose operon? 2. Describe how level of a key metabolite [tryptophane] controls the gene function using tryptophane operon
- 10. Think about the situation where there is an increased concentration of both glucose and allolactose in the media. Does this situation lead to the lac operon activation or not? Justify your answer? 11. Draw the structure of lac operon and its action when the following conditions are prevailing in the media: increased glucose, low glucose, higher c-AMP, low c-AMP, higher glucose, and high c-AMP, low glucose and High c-AMP, Higher glucose and higher allolactose? please assist with the above two questions4. Predict the level of genetic activity of the lac operon as well as the status of the lac repressor and the CAP protein under the cellular conditions listed in the following table (+ = sugar present; = sugar absent): Cellular conditions Lactose Glucose + + + + a) Operon (on or off or basal?) b) Operon (on or off or basal?) Lac repressor (bound to operator?) CAP protein (bound to CAP site?) d) Operon (on or off or basal?) PROE Lac repressor (bound to operator?) CAP protein (bound to CAP site?) c) Operon (on or off or basal?) Lac repressor (bound to operator?) CAP protein (bound to CAP site?) Lac repressor (bound to operator?) CAP protein (bound to CAP site?)1. Draw a diagram of the lac operon. PO I+P+O+Z+Y+ I+P+OcZ+Y- I-P+O+Z+Y+ Lab section (circle): lacz gene lac Y gene lacA gene I+P+O+ZY/IPO+Z+Y- 2. The promoter (P) is the start site of transcription through the binding of RNA polymerase before actual mRNA production. Mutated promoters (P) cannot bind RNA polymerase, so no mRNA is made. In the table below, make predictions about the expression of ß-galactosidase and permease for different partial diploid (F') genotypes. Do this for E. coli growth on two different types of media - one without lactose, and one with. Use a to mark where there is no enzyme produced, and a + to mark where the enzyme is made. The first genotype has all wild-type alleles, and has been filled in as an example. Genotype IPOCZ+Y/I-P+O+Z-Y+ ISP+O+Z+Y/I+P+O+Z*Y* CR 1:00 Lactose absent B-galactosidase Permease R 3:-00 + R 6:30 Lactose present B-galactosidase F 10:00 Permease
- 1. Describe which enzymes are required for lactose and tryptophan metabolism inbacteria when lactose and tryptophan, respectively, are (a) present and (b)absent. 2. Contrast positive versus negative regulation of gene expression. Describe therole of the repressor in an inducible system and in a repressible system.5. There are four genes in the FAK operon. a. How many mRNAs are encoded by this operon? b. The FAK operon has a weak promoter, where RNA polymerase does not bind well. What type of molecule could directly help RNA polymerase to bind more tightly to this promoter? i. An inducer 6. Consider the Lac operon of E. coli, which is normally expressed only when lactose is present and glucose absent. You identify a mutant strain of bacteria in which Lacz, Lacy, and LacA, are expressed in the absence of glucose, whether or not lactose is present. a. ii. An activator iii. A promoter iv. A repressor v. A ligand C. Assume that one mutation causes this mis-regulation. Which interaction must be disrupted by this mutation? i. Interaction between RNA polymerase and the lac promoter ii. Interaction between Lacl protein and the lac operator (its binding site) Interaction between CRP and CAMP Interaction between CRP-CAMP (aka CAP) its binding site adjacent to lac operon Interaction between Lacl protein…1. Utilizing lactose operon, how can we explain how induction controls enzyme synthesis 2. Use tryptophane operon and describe how levels of a key metabolite/tryptophane controls the gene function
- 1. IPTG can be used in the laboratory as a synthetic inducer of the lac operon, instead of lactose. Using your knowledge of how the lac operon is regulated, predict whether these gene products (B -galactosidase, lactose Permease) will be produced (yes (+)/no (-)) in the presence or absence of the inducer IPTG in the following partial diploids +IPTG - ΙPTG B-galactosidase Permease B-galactosidase Permease (A) I+P-O+Z-Y+/ I-P+O°Z+Y- (B) I'P+O°Z+Y-/ I+P+O+Z-Y+ w (C) I+P+O+Z-Y+/ I-P+O+Z+Y- (D) I+P+O°Z-Y+/ I°P+O+Z+Y+ (E) I+P-O°Z+Y+/ I-P+O+Z+Y+3. Bacteria are normally haploid organisms (1 copy of the genome), however it is possible to introduce a plasmid into the bacteria to create a pseudo-diploid situation. Here we have introduced a plasmid that carries the lac operon. Based on the following genotypes, predict the state of the beta-glactosidase enzyme (lac Z) in each of the following situations. I- = a mutation in the repressor protein that cannot bind to the operator Is = a mutation in the repressor protein that cannot be removed from the operator Oc = a mutation in the operator that cannot bind the repressor Z` = a mutation in beta-galactosidase that makes the enzyme non-functional Chromosome Lactose Present I, Z, O I, Z, O I. Z-.O I-,Z,O IS,Z-,Oc, Plasmid I, Z, O I, Z, O I, Z, Oc I,Z,O I,Z,O Yes No No Yes Yes Functional enzyme (yes/no) Phenotype (inducible constitutive, repressed constitutive regressed1. Assuming that the level of glucose is low, a mutation in the repressor associated with the lac operon of E. coli which prevents binding of the repressor to lactose should result in: a. constitutive expression of the lac operon genes b. lack of expression or refuced expression of the lac operon genes under all circumstances c. expression of the genes only when lactose is present d. expression of the genes only when lactose is absent e. lack of expression of the lac operon genes, resulting in an increase in lactose production