The arabinose operon How does the araC protein regulate of the arabinose operon. How does its binding [to what] differ in the presence or absence of arabinose? How is transcription thought to be influenced?
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![The arabinose operon
How does the araC protein regulate of the arabinose operon. How does its binding [to what] differ in the presence or
absence of arabinose? How is transcription thought to be influenced?
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- You are studying an operon containing three genesthat are cotranscribed in the order hupF, hupH, andhupG. Diagram the mRNA for this operon, showingthe location of the 5′ and 3′ ends, all open readingframes, translational start sites, stop codons, transcription termination signals, and any regions thatmight be in the mRNA but do not serve any of thesefunctions.. In an effort to determine the location of an operator sitefor a negatively regulated gene, you have made a seriesof deletions within the regulatory region. The extent ofeach deletion is shown by the line underneath the sequence, and the resulting expression from the operon(i = inducible; c = constitutive; − = no expression) isalso indicated.... GGAT C T T AGCCGGCTAACATGATAAATATAA......C C T AGAATCGGCCGA TTGTA C T A TTT ATAT T ...1 i2 –3 c4 –5 ca. What can you conclude from these data about thelocation of the operator site?b. Why do you think deletions 2 and 4 show no expression of the gene?One of the main lessons of this chapter is that severalbacterial genes are often transcribed from a singlepromoter into a large multigene (polycistronic) transcript. The region of DNA containing the set of genesthat are cotranscribed, along with all of the regulatoryelements that control the expression of these genes, iscalled an operon.a. Which of the mechanisms in the following listcould explain differences in the levels of themRNAs for different operons?b. Which of the mechanisms in the following listcould explain differences in the levels of theprotein products of different genes in the sameoperon?i. Different promoters might have different DNAsequences.ii. Different promoters might be recognized bydifferent types of RNA polymerase.iii. The secondary structures of mRNAs might differ so as to influence the rate at which they aredegraded by ribonucleases.PROBLEMSProblems 575a. Why did the mating with a nonlysogenic recipientresult in phage growth and release, but the infectionof a…
- A mutant strain has a defective lac operator site that results in theconstitutive expression of the lac operon. Outline an experimentyou would carry out to demonstrate that the operator site mustbe physically adjacent to the genes that it influences. Based onyour knowledge of the lac operon, describe the results you wouldexpect.The symbols a, b, and c in the table below represent theE. coli lac system genes for the repressor (I ), the operator(O) region, and the β-galactosidase (Z), although notnecessarily in that order. Furthermore, the order inwhich the symbols are written in the genotypes is notnecessarily the actual sequence in the lac operon.Activity (+) or inactivity (−) of Z geneGenotype Inducer absent Inducer presenta- b+ c+ + +a+ b+ c- + +a+ b- c- - a+ b- c+/a- b+ c- + +a+ b+ c+/a- b- c- - +a+ b+ c-/a- b- c+ - +a- b+ c+/a+ b- c- + +a. Which symbol (a, b, or c) represents each of the lacgenes I, O, and Z ?b. In the table, a superscript minus sign on a gene symbolmerely indicates a mutant, but some mutant behaviorsin this system are given special mutant designations.Using the conventional gene symbols for the lac operon,designate each genotype in the table.Six strains of E. coli (mutants 1–6) that had one of thefollowing mutations (i–vi) affecting the lac operonwere isolated.i. deletion of lacYii. ocmutationiii. missense mutation in lacZiv. inversion of the lac operon (but not an inversion ofthe lacI gene)v. superrepressor mutationvi. inversion of lacZ, Y, and A but not lacI, P, oa. Which of these mutations would prevent the strainfrom utilizing lactose?b. The entire lac operon (including the lacI gene andits promoter) from each of the six E. coli strainswas cloned into a plasmid vector containing an ampicillin resistance gene. Each recombinant plasmidwas transformed into each of the six strains to create partial diploids. In analysis of these strains, mutant 1 was found to carry a deletion of lacY, so thisstrain corresponds to mutation i in the list above.Which of the other types of mutations would be expected to complement mutant 1 in these partial diploids so as to allow lactose utilization?c. In part (b), each strain was plated on…
- Suppose that you have isolated a mutant strain of E. coli inwhich the lac operon is constitutively expressed. To understandthe nature of this defect, you create a merozygote in which themutant strain contains an F′ factor with a normal lac operonand a normal lacI gene. You then compare the mutant strain andthe merozygote with regard to their β-galactosidase activities inthe presence and absence of lactose. You obtain the followingresults:Please help me iwht this question explain how the regulation of the Lac Operon incorporate both negative and positive regulation?The promoter of an operon is the site to which RNApolymerase binds to begin transcription. Certain basechanges in the promoter result in a mutant site towhich RNA polymerase cannot bind. Would you expect mutations in the promoter that prevent binding ofRNA polymerase to act in trans on another copy ofthe operon on a plasmid in the cell, or only in cis onthe copy immediately adjacent to the mutated site?
- You have isolated two different mutants (reg1 andreg2) causing constitutive expression of the emu operon (emu1 emu2). One mutant contains a defect in aDNA-binding site, and the other has a loss-of-functiondefect in the gene encoding a protein that binds tothe site.a. Is the DNA-binding protein a positive or negativeregulator of gene expression?b. To determine which mutant has a defect in the siteand which one has a mutation in the binding protein, you decide to do an analysis using F′ plasmids. Assuming you can assay levels of the Emu1and Emu2 proteins, what results do you predict forthe two strains (i and ii; see descriptions below) ifreg2 encodes the regulatory protein and reg1 is theregulatory site?i. F′ (reg1−reg2+emu1−emu2+)/reg1+reg2+emu1+emu2−ii. F′ (reg1+reg2−emu1−emu2+)/reg1+reg2+emu1+emu2−c. What results do you predict for the two strains(i and ii) if reg1 encodes the regulatory proteinand reg2 is the regulatory site?F’ lac I- O+ Z+/ lac I+ Oc Z- the genotype indicated would lead to what type of expression of β‐galactosidase with respect to lactose inducibility of the lac operon. A. Constitutive B. Cannot be determined C. Inducible D. Uninducible40The accepted term for the functional investigation of a gene that starts with the gene sequence rather than a mutant phenotype is forward genetics. Yesorno 41Alu elements in the human genomes are examples of SINE elements. Yesorno 42The lac operon has three lac operators. Yesorno