BIOLOGY
12th Edition
ISBN: 9781260169614
Author: Raven
Publisher: RENT MCG
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Chapter 16, Problem 1S
You have isolated a series of mutants affecting regulation of the lac operon. All of these are constitutive, that is, they express the lac operon all the time. You also have both mutant and wild-type alleles for each mutant in all combinations, and on F′ plasmids, which can be introduced into cells to make the cell diploid for the relevant genes. How would you use these tools to determine which mutants affect DNA binding sites on DNA, and which affect proteins that bind to DNA?
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Suppose you have six strains of E. coli. One is wildtype, and each of the other five has a single one of thefollowing mutations: lacZ−, lacY−, lacI−, oc, andlacIS. For each of these six strains, describe thephenotype you would observe using the following assays. [Notes: (1) IPTG is a colorless synthetic molecule that acts as an inducer of lac operon expressionbut cannot serve as a carbon source for bacterialgrowth because it cannot be cleaved byβ-galactosidase; (2) X-gal cannot serve as a carbonsource for growth; (3) E. coli requires active lactosepermease (the product of lacY) to allow lactose,X-gal, or IPTG into the cells.] Colony color in medium containing glycerol as theonly carbon source and X-gal, but no IPTG.d. Colony color in medium containing high levels ofglucose as the only carbon source, X-gal, andIPTG.e. Colony color in medium containing high levels ofglucose as the only carbon source and X-gal, butno IPTG
. Suppose you have six strains of E. coli. One is wildtype, and each of the other five has a single one of thefollowing mutations: lacZ−, lacY−, lacI−, oc, andlacIS. For each of these six strains, describe thephenotype you would observe using the following assays. [Notes: (1) IPTG is a colorless synthetic molecule that acts as an inducer of lac operon expressionbut cannot serve as a carbon source for bacterialgrowth because it cannot be cleaved byβ-galactosidase; (2) X-gal cannot serve as a carbonsource for growth; (3) E. coli requires active lactosepermease (the product of lacY) to allow lactose,X-gal, or IPTG into the cells.]a. Growth on medium in which the only carbonsource was lactose.b. Colony color in medium containing glycerol as theonly carbon source, X-gal, and IPTG
A mutant strain of E. coli has a premature stop mutation in the lacZgene, resulting in a non-functional b-galactosidase. Otherwise, all other parts of the operon are functional. Which component of an F' plasmid will restore normal regulation and function of the lac operon in the resulting partial diploid?
Chapter 16 Solutions
BIOLOGY
Ch. 16.1 - Prob. 1LOCh. 16.1 - Prob. 2LOCh. 16.1 - Prob. 3LOCh. 16.2 - Explain how proteins can interact with base-pairs...Ch. 16.2 - Prob. 2LOCh. 16.3 - Prob. 1LOCh. 16.3 - Prob. 2LOCh. 16.3 - Explain control of gene expression in the trp...Ch. 16.4 - Prob. 1LOCh. 16.4 - Prob. 2LO
Ch. 16.4 - Prob. 3LOCh. 16.5 - Describe at least two kinds of epigenetic mark.Ch. 16.5 - Explain the function of chromatin-remodeling...Ch. 16.6 - Prob. 1LOCh. 16.6 - Prob. 2LOCh. 16.7 - Prob. 1LOCh. 16.7 - Prob. 2LOCh. 16 - Prob. 1DACh. 16 - What advantage might a bacterium gain by linking...Ch. 16 - Prob. 2IQCh. 16 - Prob. 3IQCh. 16 - In prokaryotes, control of gene expression usually...Ch. 16 - Prob. 2UCh. 16 - Prob. 3UCh. 16 - The lac operon is controlled by two main proteins....Ch. 16 - In eukaryotes, binding of RNA polymerase to a...Ch. 16 - In eukaryotes, the regulation of gene expression...Ch. 16 - In the trp operon, the repressor binds to DNA a....Ch. 16 - Prob. 1ACh. 16 - Specific transcription factors in eukaryotes...Ch. 16 - Repression in the trp operon and induction in the...Ch. 16 - Regulation by small RNAs and alternative splicing...Ch. 16 - Eukaryotic mRNAs differ from prokaryotic mRNAs in...Ch. 16 - In the cell cycle, cyclin proteins are produced in...Ch. 16 - A mechanism of control in E. coli not discussed in...Ch. 16 - You have isolated a series of mutants affecting...Ch. 16 - Examples of positive and negative control of...Ch. 16 - What forms of eukaryotic control of gene...Ch. 16 - The number and type of proteins found in a cell...
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- 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_forwardIf a wild-type (normal, NOTmutated) E. coli strain is grown in a medium: a. without lactose or glucose, how many proteins (and which ones) are bound to the lac operon? b. Without lactose, but with glucose, how many proteins (and which ones) are bound to the lac operon??arrow_forwardThe following shows the genotype of a partial diploid bacterial cell - where one chromosomal region containing the lac operon in E,coli is given, and the other fragment is from a plasmid carrying another lac operon from another source. The two are separated by a slash (/). The possible answers indicate with a ʺ+ʺ or a ʺ-ʺ whether β-galactosidase would be expected to be produced at induced levels under two circumstances: 1) first in the absence of lactose and 2) second in the presence of lactose. (Assume that glucose is not present in the medium.)Genotype F: I+ Oc Z-/ Fʹ I- O+ Z+ KEY:I+ = wild-type repressorI- = mutant repressor (unable to bind to the operator)Is = mutant repressor (insensitive to lactose)O+ = wild-type operatorOc = constitutive operator (insensitive to repressor)arrow_forward
- Give all possible genotypes of a lac operon that produces, or fails to produce, β-galactosidase and permease under the following conditions. Do not give partial-diploid genotypes. Lactose absent Lactose present β-Galactosidase Permease β-Galactosidase Permease a. − − + + b. − − − + c. − − + − d. + + + + e. − − − − f. + − + − g. − + − +arrow_forwardWhat 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.arrow_forwardYou then make a screen to identify potential mutants (shown as * in the diagram) that are able to constitutively activate Up Late operon in the absence of Red Bull and those that are not able to facilitate E. Coli growth even when fed Red Bull. You find that each class of mutations localize separately to two separate regions. For those mutations that prevent growth even when fed Red Bull are all clustered upstream of the core promoter around -50 bp. For those mutations that are able to constitutively activate the operon in the absence of Red Bull are all located between the coding region of sleep and wings. Further analysis of each DNA sequence shows that the sequence upstream of the promoter binds the protein wings and the region between the coding sequence of sleep and wings binds the protein sleep. When the DNA sequence of each is mutated, the ability to bind DNA is lost. Propose a final method of gene regulation of the Up Lateoperon using an updated drawn figure of the Up Late…arrow_forward
- A researcher engineers a lac (lactose) operon on a plasmid but inactivates all parts of the lac operator (lacO) and the lac promoter, replacing them with the binding site for the LexA repressor (which acts in SOS response) and a promoter regulated by LexA. The plasmid is introduced into E. coli cells that have a lac operon with an inactive lacZ gene. Under what conditions will these transformed cells produce beta-galactosidase?arrow_forwardYou 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. Is the DNA-binding protein a positive or negative regulator of gene expression?arrow_forwardLet’s suppose you have isolated a mutant strain of E. coli in which the lac operon is constitutively expressed. In other words, the operon is turned on in the presence or absence of lactose. One possibility is that the mutation may block the transcription of the lacI gene, thereby preventing the synthesis of lac repressor. A second possibility is that the mutation could alter the sequence of the lac operon in a way that prevents the repressor protein from binding to the operator. How would you distinguish between these two possibilities?arrow_forward
- In addition to observing similarities to the lac operon, you also notice that this gene is regulated via attenuation, similar to the trp operon. Based on this similarity to this model operon, you could state that ___________. Group of answer choices If a terminator loop forms in the DNA, the expression of the structural genes is halted. The formation of the terminator hairpin followed by a series of Uracil (UUUUUUU) functions similar to Rho-Independent termination to result in the stopping of transcription. The transcription of a leader sequence affects the translation of the structural genes. Never mind – all of these statements are true! Attenuation will be the primary means of transcriptional regulation, with a repressor used as a back up option.arrow_forwardYou 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_forwardMany amino acid biosynthetic operon under attenuation control are also under negative control. Considering that the environment of a bacterium can be highly dynamic, what advantage could be conferred by having attenuation as a second layer of control?arrow_forward
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