6. The transposon Tn5 was used to create a library of 50,000 Escherichia coli mutant strains...
6. The transposon Tn5 was used to create a library of 50,000 Escherichia coli mutant strains having random Tn5 insertions in their chromosomal genes. A suicide delivery plasmid vector that carries Tn5 was used for this purpose. Explain how and why a suicide delivery plasmid was used to generate this library and give one (1) example of how you would screen this library to find mutants with Tn5 insertions in specific genes of interest, for example, in the lac (lactose) operon. (10 marks).
Homework Answers
Suicide delivery plasmid vector does not have ori site in its
plasmid so plasmid not able to replicate in the bacterial
cell.
After number of generation the plasmid started to dilute in the
cell and it gets eliminated is refer as committing suicide. In Tn5
transposon the transposon are transfered and unwanted part of
vector get diluted. In lac operon insertion the transpon Tn5 have
same restriction enzyme sites in lac operon and same sequence
present before and after of transposon construct. After this vector
insert into bacterial cell transposon or jump gene transfer from
insert in to lac operon.
Add Answer to:
6. The transposon Tn5 was used to create a library of 50,000
Escherichia coli mutant strains...
Three different strains of E. coli carry a mutation in the lac operon and/or laci gene....
Three different strains of E. coli carry a mutation in the lac operon and/or laci gene. The production of B-galactosidase (+ present or - absent) is measured when lactose is present and absent from the medium. Assume the mutations involve only 1, 0, or Z. A merodiploid is constructed for each of the three strains. The plasmid carries a wild type lac operon and lacl gene. The production of functional B-galactosidase (+ present or - absent) is measured when lactose...
As a student project, you have isolated six new mutant strains of E. coli with altered...
As a student project, you have isolated six new mutant strains
of E. coli with altered behavior of the lactose operon. The strains
are listed in the table below, together with their phenotypes (with
regard to significant ?-galactosidase synthesis) in three specific
situations.
Columns 1 and 2 present the phenotypes of each mutant haploid
strain. In column 1, the mutant is in an otherwise wild-type
genome. In column 2, the genome also carries a nonsense-suppressor
mutation (that is not present...
Shown below are relevant genes and sites from various E. coli strains. Note: + designates the...
Shown below are relevant genes and sites from various E. coli strains. Note: + designates the wild-type gene or site that is fully functional, and - designates a deletion of that gene or site; lacot means all operators (01, O2 and 03) are functional; assume all other genes and sites not listed are wild-type and functional (including lacA). In some cases, a plasmid containing a wild-type functional gene or site was transformed into the E. coli strain. The strains were...
QUESTION 8 The following situations (1-4) involve different types of gene regulation in prokaryotic cells as shown. OFF...
QUESTION 8 The following situations (1-4) involve different types of gene regulation in prokaryotic cells as shown. OFF and ON reter to whether the gene is transcribed or not. Draw clearly-labelled regulatory proteins and effector molecules in each diagram to explain how the regulation works in each case. The first one has been done for you as an example a) (6 marks) Type of Regulation Regulatory protein effector molecule 1 DNA X negative inducible OFF ON 2 negative repressible DNA...
Shown below are relevant genes and sites from various E. coli strains. Note: + designates the...
Shown below are relevant genes and sites from various E. coli strains. Note: + designates the wild-type gene or site that is fully functional, and – designates a deletion of that gene or site; lacO+ means all operators (O1, O2 and O3) are functional; assume all other genes and sites not listed are wild-type and functional (including lacA). In some cases, a plasmid containing a wild-type functional gene or site was transformed into the E. coli strain. The strains were...
Shown below are relevant genes and sites from various E. coll strains. Note: + designates the...
Shown below are relevant genes and sites from various E. coll strains. Note: + designates the wild-type gene or site that is fully functional, and - designates a deletion of that gene or site; laco means all operators (Oj, 0, and Oy) are functional; assure all other genes and sites not listed are wild-type and functional (including locA). In some cases, a plasmid caintaining a wild-type functional gene or site was transformed into the E. coli strain. The strains were...
The following set of three separate experiments was conducted. Two strains of Escherichia coli are used....
The following set of three separate experiments was conducted. Two strains of Escherichia coli are used. Strain 1 needs histidine added to the medium to grow. Strain 2 needs methionine added to the medium to grow. Experiment 1: After strain 1 and strain 2 are mixed in a culture flask containing a minimal medium (containing no added amino acids) and allowed to grow overnight. Results: It is possible isolate new strains that grow on the minimal medium. Experiment 2: Strains...
3. (2 points) In the nematode roundworm, Caenorhabditis elegans, the LIN-14 protein controls the timing of...
3. (2 points) In the nematode roundworm, Caenorhabditis elegans, the LIN-14 protein controls the timing of certain cell divisions during development. LIN-14 protein levels are normally high in early development but decrease in the later stages. In a lin-4 mutant, the level of LIN-14 protein stays high throughout development, changing the pattern of cell divisions in the animal and producing defects in the shape of the animal. The lin-4 gene encodes a microRNA that binds to a sequence in the...
2. A dominant allele H reduces the number of body bristles that Drosophila flies have, giving...
2. A dominant allele H reduces the number of body bristles that Drosophila flies have, giving rise to a “hairless” phenotype. In the homozygous condition, H is lethal. An independently assorting dominant allele S has no effect on bristle number except in the presence of H, in which case a single dose of S suppresses the hairless phenotype, thus restoring the "hairy" phenotype. However, S also is lethal in the homozygous (S/S) condition. What ratio of hairy to hairless flies...
Three different strains of E. coli carry a mutation in the lac operon and/or laci gene. The production of B-galactosidase (+ present or - absent) is measured when lactose is present and absent from the medium. Assume the mutations involve only 1, 0, or Z. A merodiploid is constructed for each of the three strains. The plasmid carries a wild type lac operon and lacl gene. The production of functional B-galactosidase (+ present or - absent) is measured when lactose...
As a student project, you have isolated six new mutant strains
of E. coli with altered behavior of the lactose operon. The strains
are listed in the table below, together with their phenotypes (with
regard to significant ?-galactosidase synthesis) in three specific
situations.
Columns 1 and 2 present the phenotypes of each mutant haploid
strain. In column 1, the mutant is in an otherwise wild-type
genome. In column 2, the genome also carries a nonsense-suppressor
mutation (that is not present...
Shown below are relevant genes and sites from various E. coli strains. Note: + designates the wild-type gene or site that is fully functional, and - designates a deletion of that gene or site; lacot means all operators (01, O2 and 03) are functional; assume all other genes and sites not listed are wild-type and functional (including lacA). In some cases, a plasmid containing a wild-type functional gene or site was transformed into the E. coli strain. The strains were...
QUESTION 8 The following situations (1-4) involve different types of gene regulation in prokaryotic cells as shown. OFF and ON reter to whether the gene is transcribed or not. Draw clearly-labelled regulatory proteins and effector molecules in each diagram to explain how the regulation works in each case. The first one has been done for you as an example a) (6 marks) Type of Regulation Regulatory protein effector molecule 1 DNA X negative inducible OFF ON 2 negative repressible DNA...
Shown below are relevant genes and sites from various E. coll strains. Note: + designates the wild-type gene or site that is fully functional, and - designates a deletion of that gene or site; laco means all operators (Oj, 0, and Oy) are functional; assure all other genes and sites not listed are wild-type and functional (including locA). In some cases, a plasmid caintaining a wild-type functional gene or site was transformed into the E. coli strain. The strains were...
The following set of three separate experiments was conducted. Two strains of Escherichia coli are used. Strain 1 needs histidine added to the medium to grow. Strain 2 needs methionine added to the medium to grow. Experiment 1: After strain 1 and strain 2 are mixed in a culture flask containing a minimal medium (containing no added amino acids) and allowed to grow overnight. Results: It is possible isolate new strains that grow on the minimal medium. Experiment 2: Strains...
3. (2 points) In the nematode roundworm, Caenorhabditis elegans, the LIN-14 protein controls the timing of certain cell divisions during development. LIN-14 protein levels are normally high in early development but decrease in the later stages. In a lin-4 mutant, the level of LIN-14 protein stays high throughout development, changing the pattern of cell divisions in the animal and producing defects in the shape of the animal. The lin-4 gene encodes a microRNA that binds to a sequence in the...
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