Microbiology: An Evolving Science (Fourth Edition)
4th Edition
ISBN: 9780393615098
Author: John W. Foster, Joan L. Slonczewski
Publisher: W. W. Norton & Company
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Chapter 10.2, Problem 3TQ
Summary Introduction
To review:
The expression of lacZ gene when a plasmid containing mutated genes is introduced.
Introduction:
Plasmid is an extrachromosomal DNA (deoxyribonucleic acid). The plasmids are smaller in comparison to the nuclear chromosome. These entities are a characteristic feature of the bacterial cells but certain eukaryotic cells such as yeasts also contain plasmids. The plasmids generally confer antibiotic resistance to the bacterial cells.
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Given the following genotypes, explain, by answering the questions in each number, how the mutation (identified by a (-) superscript) will affect E. coli grown in lactose medium.
Will there be a complete set ofgene products? (Yes/No)
Will the lac operon be turnedon/off?
Will the cell survive? (Yes/No)
a. i + p + o + z - y +
b. i + p - o + z + y +
c. i + p + o - z + y +
O The lac operon in E.coli encodes enzymes necessary for the breakdown of lactose. For
each enzyme (lac Z and lac Y), indicate with a + or-whether or not it is made when
there is no lactose or when there is lactose.
B-galactosidase (lac Z)
No Lactose
Permease (lac Y)
Lactose
Lactose
No Lactose
Genotype
PP0 Z Y/I P*O*Z•Y*
I'POCZ Y*/I P* O©Z*Y°
P O Z'Y/I P'OʻZ'Y*
PP O ZY*/IP*O*Z*Y*
IP OCZ Y /I P*O*Z•Y*
IPO ZY*/I* P*O©Z*Y•
I'PO*Z Y*/IP'O*Z*Y°
Strain LOLZ has a mutation in the lacI gene that results in a lacI protein that cannot bind allolactose. Otherwise all other parts of the operon are functional.
Circle the least number of components for an F' plasmid that will restore normal regulation and function of the lac operon in the resulting partial diploid. (may need more than one)
(a) lacI+ (b) lacO+ (c) lacP+ (d) lacZ+ (e) None, cannot be restored.
Chapter 10 Solutions
Microbiology: An Evolving Science (Fourth Edition)
Ch. 10.1 - Prob. 1TQCh. 10.2 - Prob. 1TQCh. 10.2 - Prob. 2TQCh. 10.2 - Prob. 3TQCh. 10.2 - Prob. 4TQCh. 10.2 - Prob. 5TQCh. 10.3 - Prob. 1TQCh. 10.3 - Prob. 2TQCh. 10.4 - Prob. 1TQCh. 10.5 - Prob. 1TQ
Ch. 10.5 - Prob. 2TQCh. 10.6 - Prob. 1TQCh. 10 - Prob. 1RQCh. 10 - Prob. 2RQCh. 10 - Prob. 3RQCh. 10 - Prob. 4RQCh. 10 - Prob. 5RQCh. 10 - Prob. 6RQCh. 10 - Prob. 7RQCh. 10 - Prob. 8RQCh. 10 - Prob. 9RQCh. 10 - Prob. 10RQCh. 10 - Prob. 11RQCh. 10 - Prob. 12RQCh. 10 - Prob. 13RQCh. 10 - Prob. 14RQCh. 10 - Prob. 1TQCh. 10 - Prob. 2TQCh. 10 - Prob. 3TQCh. 10 - Prob. 4TQ
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- Liliana is preparing chemically competent cells for heat shock transformation from old batches of competent cells available in the freezers. The competent cells would eventually be used for the expression of a prokaryotic enzyme using the pET vector system. (1) (ii) Why is Escherichia coli DH5a suitable to propagate the plasmid before protein expression? During the protein expression step, the plasmids are transformed into a different host cell, BL21(DE3). Explain why this step is pertinent to express the protein of interest.arrow_forwardThe map of the lac operon is shown below. Consider the following examples that include both haploids and partial diploids and explain in each scenario whether the repressor can bind and regulate expression and 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+ Oc Z+ Y+arrow_forwardA sample of E.coli cells is transformed with a plasmid containing a transcriptional fusion including the promoter of an E.coli housekeeping gene and the LacZ gene. The transformed cells are plated out on selective medium containing X-gal. Which of the following best describes the likely results. a. Colonies will only be blue if lactose is also included in the growth medium. b. Colonies will only be blue if lactose and glucose are included in the growth medium c. Most of the colonies should turn blue. d. Presence of any white colonies may indicate deletion or mutation of the promoter. e. Both c. and d. are correctarrow_forward
- For each of the E. coli strains containing lac operon alleles listed, indicate whether the strain is inducible, constitutive, or unable to express beta-galactosidase and permease. (P+ and P- are functional and nonfunctional promoters, respectively) I+ P+ o+ Z- Y+ / I+ P+ oc Z+ Y+ I+ P+ o+ Z+ Y+ / I- P+ oc Z+ Y- I+ P+ o+ Z- Y+ / I- P+ oc Z+ Y- I- P- o+ Z+ Y- / I+ P+ oc Z- Y+ IS P+ o+ Z+ Y+ / I- P+ o+ Z+ Y-arrow_forwardStrain 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. Circle the least number of components for an F' plasmid that will restore normal regulation and function of the lacoperon in the resulting partial diploid. (may need more than one) (a) lacI+ (b) lacO+ (c) lacP+ (d) lacZ+ (e) None, cannot be restored.arrow_forwardA number of mutations affect the expression of the lac operon in E. coli. The genotypes of several E. coli strains are shown below. ("+" indicates a wild-type gene with normal function and "-" indicates a loss-of-function allele.) Please predict which of the following strains would have the highest beta-galactosidase enzyme activity, when grown in the lactose medium. O CAP+ r* p* o* z O CAP* I P* o* z* O CAP* r* P O* z* O CAP I P* O z*arrow_forward
- The 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_forwardStrain X-23 has a mutation in the lacI gene resulting in a lacI protein that cannot bind DNA. Otherwise all other parts of the operon are functional. Circle the least number of components for an F' plasmid that will restore normal regulation and function of the lacoperon in the resulting partial diploid. (may need more than one) (a) lacI+ (b) lacO+ (c) lacP+ (d) lacZ+ (e) None, cannot be restoredarrow_forwardA number of mutations affect the expression of the lac operon in E. coli. The genotypes of several E. coli strains are shown below. ("+" indicates a wild-type gene with normal function and "-" indicates a loss-of-function allele.) Please predict which of the following strains would have the lowest beta-galactosidase enzyme activity, when grown in the lactose medium. OF POZY Ort Ptot Z¹ Yt Ort p²o+z¹Y+ Orpt ot ztyarrow_forward
- Based on the following wild type DNA sequence, indicate if each of the mutations should be classified as : insertion, deletion, missense, nonsense, silent (Use the provided Genetic Code table and remember you have been given DNA sequence). Wild Type: 5’ ATG GCT AGA GTC GAG TTG 3’ Mutant 1: 5’ ATG GCA GAG TCG AGT TG 3’ Mutant 2: 5’ ATG GCT TGA GTC GAG TTG 3’ Mutant 3: 5’ ATG GCT AGA GTT GAG TTG 3’ Mutant 4: 5’ ATG GCT AGA AGT CGA GTT G 3’ Mutant 5: 5’ ATG GCT AGA ATC GAG GTT 3’arrow_forwardWe are utilizing BL21 DE3 bacterial cells for the expression of the ADA protein via autoinduction. Create a schematic/figure showing the biological mechanism for expressing our desired protein in this cell line. We use the DE3 lysogen for expressing T7 polymerase and our plasmid has kanamycin resistance (not ampicillin).Create a schematic of this expression.arrow_forwardA number of mutations affect the expression of the lac operon in E. coli. The genotypes of several E. coli strains are shown below. ("+" indicates a wild-type gene with normal function and "-" indicates a loss-of-function allele.) Please predict which of the following strains would have the lowest beta-galactosidase enzyme activity, when grown in the lactose medium. Orpt o* z* r* Orpt ot z* Y OrptoztY Orrotzr OrPotz*Yarrow_forward
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