ND STONY BROOK UNIVERSITY LOOSELEAF GENETICS: FROM GENES TO GENOMES
6th Edition
ISBN: 9781260406092
Author: HARTWELL, Leland, HOOD, Leroy, Goldberg, Michael
Publisher: Mcgraw-hill Education/stony Brook University
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Chapter 16, Problem 11P
Mutants were isolated in which the constitutive
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The logic of the Lac operon was deciphered by using bacterial lac-operon mutants. Crucial to the experiments was creation of partial diploid cells in which operon from a wild-type cell is introduced into the mutant cell [thus there are 2 copies of the operon], and its ability to restore repression is assessed.
Three classes of mutants were found that affect expression of the lac operon. Two of these resulted in constitutive expression of the operon, and the other eliminated all expression, even in the presence of lactose.
Use the information above to complete the following table indicating with a (Y) yes or (N= No) if enzymatic activity of B-Gal will be detected (protein made and functional). Assume there is no glucose.
Remember: Promoter for operon controls ALL genes downstream of it, coding sequence of the genes themselves matter (so if transcription is occurring but structural gene is producing a non-functional protein there will be no activity detected)
For the lac operon…
A number of mutations affect the expression of the lac operon in E. coli. Consider each genotype below and complete the table using “+” to indicate that the gene is expressed, and “−” to indicate that gene is not expressed.
What experimental results would indicate that the mutation lacISlacIS is dominant to lacI+lacI+?
In lacISlacIS/lacI+lacI+ partial diploids, the lac operon is in a repressed state in the absence of lactose.
In lacISlacIS/lacI+lacI+ partial diploids, the lac operon is in a constitutive state in the absence of the repressor.
In lacISlacIS/lacI+lacI+ partial diploids, the lac operon is in an activated state in the presence of lactose.
In lacISlacIS/lacI+lacI+ partial diploids, the lac operon is in a repressed state in the absence of the repressor.
In lacISlacIS/lacI+lacI+ partial diploids, the lac operon is in a repressed state in the presence of lactose.
Chapter 16 Solutions
ND STONY BROOK UNIVERSITY LOOSELEAF GENETICS: FROM GENES TO GENOMES
Ch. 16 - For each of the terms in the left column, choose...Ch. 16 - The following statement occurs early in this...Ch. 16 - One of the main lessons of this chapter is that...Ch. 16 - All mutations that abolish function of the Rho...Ch. 16 - The figure at the beginning of this chapter shows...Ch. 16 - The promoter of an operon is the site to which RNA...Ch. 16 - You are studying an operon containing three genes...Ch. 16 - You have isolated a protein that binds to DNA in...Ch. 16 - You have isolated two different mutants reg1 and...Ch. 16 - Bacteriophage , after infecting a cell, can...
Ch. 16 - Mutants were isolated in which the constitutive...Ch. 16 - Suppose you have six strains of E. coli. One is...Ch. 16 - The previous problem raises some interesting...Ch. 16 - For each of the E. coli strains containing the lac...Ch. 16 - For each of the following growth conditions, what...Ch. 16 - For each of the following mutant E. coli strains,...Ch. 16 - Maltose utilization in E. coli requires the...Ch. 16 - Seven E. coli mutants were isolated. The activity...Ch. 16 - Cells containing missense mutations in the crp...Ch. 16 - Six strains of E.coli mutants 16 that had one of...Ch. 16 - a. The original constitutive operator mutations in...Ch. 16 - In an effort to determine the location of an...Ch. 16 - Prob. 23PCh. 16 - The footprinting experiment described in Fig....Ch. 16 - Why is the trp attenuation mechanism unique to...Ch. 16 - a. How many ribosomes are required at a minimum...Ch. 16 - The following is a sequence of the leader region...Ch. 16 - For each of the E. coli strains that follow,...Ch. 16 - Prob. 29PCh. 16 - For each element in the list that follows,...Ch. 16 - Among the structurally simplest riboswitches are...Ch. 16 - Great variation exists in the mechanisms by which...Ch. 16 - Many genes whose expression is turned on by DNA...Ch. 16 - In 2005, Frederick Blattner and his colleagues...Ch. 16 - The E.coli MalT protein is a positive regulator of...Ch. 16 - Prob. 36PCh. 16 - Prob. 37PCh. 16 - Prob. 38PCh. 16 - Prob. 39PCh. 16 - Prob. 40PCh. 16 - Prob. 41PCh. 16 - The researchers who investigated bioluminescence...Ch. 16 - Prob. 43PCh. 16 - Quorum sensing controls the expression of...Ch. 16 - Scientists are currently screening a chemical...
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- . Mutants were isolated in which the constitutive phenotype of a missense lacI mutation was suppressed.That is, the operon was now inducible. These suppressor mutations mapped to the operon, not to the lacIgene. What could these mutations be?arrow_forwardThe lac operon consists of three structural genes, lacZ, lacY and lacA that are transcribed as a single polycistronic mRNA. You are given a new strain of Escherichia coli with the following lac operon genotype: p+0°Z•Y*A +// P*O*Z*Y+ A- (i) Explain how the lac I gene affects gene expression. (ii) Explain the function of the lacP in the bacterial operon. (iii) Which part of the lac operon is cis-dominant? Explain.arrow_forwardA theoretical operon (theo) in E. coli contains several structural genes encoding enzymes that are involved sequentially in the biosynthesis of an amino acid. Unlike the lac operon, in which the repressor gene is separate from the operon, the gene encoding the regulator molecule is contained within the theo operon. When the end product (the amino acid) is present, it combines with the regulator molecule, and this complex binds to the operator, repressing the operon. In the absence of the amino acid, the regulatory molecule fails to bind to the operator, and transcription proceeds. Characterize this operon, then consider the following mutations, as well as the situation in which the wild-type gene is present along with the mutantgene in partially diploid cells (F¿):(a) Mutation in the operator region.(b) Mutation in the promoter region.(c) Mutation in the regulator gene.In each case, will the operon be active or inactive in transcription, assuming that the mutation affects the…arrow_forward
- You have isolated two different mutants (reg1 and reg2) causing constitutive expression of the emu operon (emu1 emu2). One mutant contains a defect in a DNA-binding site, and the other has a loss-of-function defect in the gene encoding a protein that binds to the site. Is the DNA-binding protein a positive or negative regulator of gene expression? Explain. To determine which mutant has a defect in the site and 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 Emu1 and Emu2 proteins, what results do you predict for the two strains (i and ii; see descriptions below) if reg2 encodes the regulatory protein and reg1 is the regulatory site? Explain. F′ (reg1− reg2+ emu1− emu2+)/reg1+ reg2+ emu1+ emu2− F′ (reg1+ reg2− emu1− emu2+)/reg1+ reg2+ emu1+ emu2−arrow_forwardTo study the lac operon, you engineer a strain of E coli to have a lac operon in which the lac Z gene is replaced by the gene for green fluorescent protein (GFP). Expression of GFP generates a green color in the cells that can be easily quantitated with a fluorescence microscope. You test the activity of the operon in the absence of the inducer IPTG, the presence of the inducer IPTG and the presence of an antibiotic the completely inhibits RNA polymerase (i.e. no gene expression). You then use this system to test the effects of various mutation on the activity of the operon. Match the following mutations with the activity (A, B or C) you would expect to observe with the mutation. All experiments are done in the presence of IPTG unless otherwise stated.arrow_forwardA mutant E. coli strain is isolated which does not ever produce the enzymes β-galactosidase or permease, irrespective of whether lactose is present or absent. If the normal wild-type genotype with respect to the lac operon can be represented as lacI+ lacP+ lacO+ lacZ+ lacY+, give three different mutant genotypes (list all 5 operon components in each case, do not consider partial diploids) which would result in the mutant phenotype described here.arrow_forward
- You have isolated different mutants (reg1 and reg2) causing constitutive expression of the emu operon (which has genes emu1 and emu2). One mutant contains a defect in a DNA-binding site, and the other has a loss-of-function defect in the gene encoding a protein that binds to the site Say you don’t know which mutant has a defect in the site and which one has a mutation in the binding protein. To figure it out, you construct the two partial diploid strains (i and ii below), and you then assay the levels of the Emu1 and Emu2 proteins in these two strains. F’ (reg1- reg2+ emu1- emu2+) / reg1+ reg2+ emu1+ emu2- F’ (reg1+ reg2- emu1- emu2+) / reg1+ reg2+ emu1+ emu2- What proteins do you predict will be expressed for strains i and ii if reg2 encodes the regulatory protein and reg1 is the regulatory site?arrow_forwardA researcher was trying to determine whether two molecules (A and B) were corepressors or inducers in their respective operon systems. Data was collected regarding the levels of protein and the amount of gene transcription for the genes in their respectiver operons. The data is shown below. Level of protein Transcription of gene 1 Transcription of gene 2 low Molecule A High low Low high high Molecule B High high high Low low low a) Determine whether molecule A and Molecule B are inducers or corepressors. Justify your answer. b) Identify which system resembles the lac operon system and which resembles the trp operon system.arrow_forwardWhat would happen if the operator sequence of the lac operon contained a mutation that prevented the repressor protein from binding the operator? (Explain what would happen both in the presence and absence of lactose)arrow_forward
- Strain P77 has a mutation in the lacO that prevents it from being bound by the lacI protein. Otherwise all other parts of the operon are functional. 1. Can strain P77 metabolize lactose if it is present? 2. If lactose is absent, will strain P77 transcribe its lac operon? 3. If lactose is present, will strain P77 transcribe its lac operon?arrow_forwardStrain ROFL4 has a premature stop mutation in the lacZ gene, resulting in a nonfunctional b-galactosidase. Otherwise all other parts of the operon are functional. 1. Can strain ROFL4 metabolize lactose if it is present? 2. If lactose is absent, will strain ROFL4 transcribe its lac operon? 3. If lactose is present, will strain ROFL4 transcribe its lac operon?arrow_forwardThe streptolysin S toxin made by S. pyogenes is encoded by a 9-gene operon, sagABCDEFGHI. Thinking about what a 3-line diagram would look like for this operon, answer the following questions. Write numeric answers only. For example, if your answer is 6 promoters, write only 6. 1) How many promoters control the expression of these genes? 2) How many locations does RNA Polymerase bind to get full expression of these genes? 3) How many ribosome binding sites are needed for full protein expression? 4) How many start codons will be needed for full protein expression? 5) How many mRNA strands will be produced with full operon expression? 6) How many proteins will be produced with full protein expression? 1arrow_forward
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