A plasmid that is both ampicillin and tetracycline resistant is cleaved with Pst l, which cleaves within the ampicillin resistance gene. The cut plasmid is ligated with Pst l-digested Drosophila DNA to prepare a genomic library, and the mixture is used to transform E. coli K12. (a) Which antibiotic should be added to the medium to select cells that have incorporated a plasmid? (b) If recombinant cells were plated on medium containing ampicillin or tetracycline and medium with both antibiotics, on which plates would you expect to see growth of bacteria containing plasmids with Drosophila DNA inserts? (c) How can you explain the presence of colonies that are resistant to both antibiotics?

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Answer 1

Textbook Question

Chapter 20, Problem 1NST

A plasmid that is both ampicillin and tetracycline resistant is cleaved with Pstl, which cleaves within the ampicillin resistance gene. The cut plasmid is ligated with Pstl-digested Drosophila DNA to prepare a genomic library, and the mixture is used to transform E. coli K12.

Chapter 20, Problem 1NST, A plasmid that is both ampicillin and tetracycline resistant is cleaved with Pstl, which cleaves

(a) Which antibiotic should be added to the medium to select cells that have incorporated a plasmid?
(b) If recombinant cells were plated on medium containing ampicillin or tetracycline and medium with both antibiotics, on which plates would you expect to see growth of bacteria containing plasmids with Drosophila DNA inserts?
(c) How can you explain the presence of colonies that are resistant to both antibiotics?

(a)

Expert Solution

Summary Introduction

To determine: The antibiotic that an individual should add to the medium to select cells that contain the recombinant plasmid.

Introduction: Plasmid refers to an circular, extrachromosomal DNA (deoxyribonucleic acid) molecule that replicates chromosome independently.

Explanation of Solution

The gene that confers resistance to tetracycline is intact in the recombinant plasmid. An individual would add tetracycline to the medium to select the recombinant plasmd in the cells. In the medium, the bacteria that have been transformed with recombinant plasmid will be resistant to tetracycline.

Thus, an individual would use tetracycline to select cells that contain the recombinant plasmid.

(b)

Expert Solution

Summary Introduction

To determine: The plates on which an individual might see the growth of bacteria having plasmids with Drosophila DNA inserts.

Introduction: Genetically modified bacterial plasmids were the first developed vectors, used for cloning purpose.

Explanation of Solution

Cloning is a important process in which bacterial cells containing recombinant DNA can be readily identified. This process is also accomplished is through the use of selectable marker genes. Antibiotic resistant genes provide very effective selectable marker genes. Therefore, the colonies that grow on tetracycline medium but do not grow on ampicillin medium would contain the Drosophila DNA insert.

Thus, colonies that only grow in tetracycline medium probably contain the Drosophila DNA insert.

(c)

Expert Solution

Summary Introduction

To determine: The reason that some colonies can grow on tetracycline medium as well as ampicillin medium.

Introduction: Plasmids have an origin of replication (ori) site, which makes it possible to produce several hundred copies of a plasmid in a single host cell.

Explanation of Solution

When performing cloning process, it is not necessary for incorporation of all DNA plasmids need to be cloned. A plasmid cut with a restriction enzyme generating sticky ends can self-ligate if cut ends of the plasmid rejoin. Also, if cleavage with PstI was incomplete, then no change in biological characteristics of the uncut plasmid would be expected.

Thus, self-ligation of plasmid and incomplete cleavage with PstI are responsible for the growth of colonies that are resistant to both antibiotics.

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Students have asked these similar questions

A plasmid that is both ampicillin and tetracyclineresistant is cleaved with PstI, which cleaves within theampicillin resistance gene. The cut plasmid is ligated withPstI-digested Drosophila DNA to prepare a genomic library,and the mixture is used to transform E. coli K12. Question: Which antibiotic should be added to the mediumto select cells that have incorporated a plasmid?

Knowing that you are using HindIII and EcoRI to cut your plasmids, and that those two enzymes cut within the MCS, use the map of pUC19 provided below to compute: What will be the sizes of the 2 restriction fragments if NO insert is present in pUC19? What will be the sizes of the 2 restriction fragments if the approximately 317 bp RT-PCR product (insert from WT satC dimer) was ligated successfully into the SmaI site? What will be the sizes of the 2 restriction fragments if TWO approximately 317 bp RT-PCR products (2 ligated inserts from WT satC dimer) were ligated into the SmaI site?

If you use the pUC18 vector to clone in the MCS region, predict the following: a) Do bacteria that are blue in color have a cloned insert? b)Do bacteria that are white in color have a cloned insert? c) If you were to grow these cells on Chloramphenicol (an antibiotic), would the bacteria with the pUC plasmid grow? Why or Why not?

Answer 2

Textbook Question

Chapter 20, Problem 1NST

A plasmid that is both ampicillin and tetracycline resistant is cleaved with Pstl, which cleaves within the ampicillin resistance gene. The cut plasmid is ligated with Pstl-digested Drosophila DNA to prepare a genomic library, and the mixture is used to transform E. coli K12.

Chapter 20, Problem 1NST, A plasmid that is both ampicillin and tetracycline resistant is cleaved with Pstl, which cleaves

(a) Which antibiotic should be added to the medium to select cells that have incorporated a plasmid?
(b) If recombinant cells were plated on medium containing ampicillin or tetracycline and medium with both antibiotics, on which plates would you expect to see growth of bacteria containing plasmids with Drosophila DNA inserts?
(c) How can you explain the presence of colonies that are resistant to both antibiotics?

(a)

Expert Solution

Summary Introduction

To determine: The antibiotic that an individual should add to the medium to select cells that contain the recombinant plasmid.

Introduction: Plasmid refers to an circular, extrachromosomal DNA (deoxyribonucleic acid) molecule that replicates chromosome independently.

Explanation of Solution

The gene that confers resistance to tetracycline is intact in the recombinant plasmid. An individual would add tetracycline to the medium to select the recombinant plasmd in the cells. In the medium, the bacteria that have been transformed with recombinant plasmid will be resistant to tetracycline.

Thus, an individual would use tetracycline to select cells that contain the recombinant plasmid.

(b)

Expert Solution

Summary Introduction

To determine: The plates on which an individual might see the growth of bacteria having plasmids with Drosophila DNA inserts.

Introduction: Genetically modified bacterial plasmids were the first developed vectors, used for cloning purpose.

Explanation of Solution

Cloning is a important process in which bacterial cells containing recombinant DNA can be readily identified. This process is also accomplished is through the use of selectable marker genes. Antibiotic resistant genes provide very effective selectable marker genes. Therefore, the colonies that grow on tetracycline medium but do not grow on ampicillin medium would contain the Drosophila DNA insert.

Thus, colonies that only grow in tetracycline medium probably contain the Drosophila DNA insert.

(c)

Expert Solution

Summary Introduction

To determine: The reason that some colonies can grow on tetracycline medium as well as ampicillin medium.

Introduction: Plasmids have an origin of replication (ori) site, which makes it possible to produce several hundred copies of a plasmid in a single host cell.

Explanation of Solution

When performing cloning process, it is not necessary for incorporation of all DNA plasmids need to be cloned. A plasmid cut with a restriction enzyme generating sticky ends can self-ligate if cut ends of the plasmid rejoin. Also, if cleavage with PstI was incomplete, then no change in biological characteristics of the uncut plasmid would be expected.

Thus, self-ligation of plasmid and incomplete cleavage with PstI are responsible for the growth of colonies that are resistant to both antibiotics.

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Answer 3

Textbook Question

Chapter 20, Problem 1NST

A plasmid that is both ampicillin and tetracycline resistant is cleaved with Pstl, which cleaves within the ampicillin resistance gene. The cut plasmid is ligated with Pstl-digested Drosophila DNA to prepare a genomic library, and the mixture is used to transform E. coli K12.

Chapter 20, Problem 1NST, A plasmid that is both ampicillin and tetracycline resistant is cleaved with Pstl, which cleaves

(a) Which antibiotic should be added to the medium to select cells that have incorporated a plasmid?
(b) If recombinant cells were plated on medium containing ampicillin or tetracycline and medium with both antibiotics, on which plates would you expect to see growth of bacteria containing plasmids with Drosophila DNA inserts?
(c) How can you explain the presence of colonies that are resistant to both antibiotics?

(a)

Expert Solution

Summary Introduction

To determine: The antibiotic that an individual should add to the medium to select cells that contain the recombinant plasmid.

Introduction: Plasmid refers to an circular, extrachromosomal DNA (deoxyribonucleic acid) molecule that replicates chromosome independently.

Explanation of Solution

The gene that confers resistance to tetracycline is intact in the recombinant plasmid. An individual would add tetracycline to the medium to select the recombinant plasmd in the cells. In the medium, the bacteria that have been transformed with recombinant plasmid will be resistant to tetracycline.

Thus, an individual would use tetracycline to select cells that contain the recombinant plasmid.

(b)

Expert Solution

Summary Introduction

To determine: The plates on which an individual might see the growth of bacteria having plasmids with Drosophila DNA inserts.

Introduction: Genetically modified bacterial plasmids were the first developed vectors, used for cloning purpose.

Explanation of Solution

Cloning is a important process in which bacterial cells containing recombinant DNA can be readily identified. This process is also accomplished is through the use of selectable marker genes. Antibiotic resistant genes provide very effective selectable marker genes. Therefore, the colonies that grow on tetracycline medium but do not grow on ampicillin medium would contain the Drosophila DNA insert.

Thus, colonies that only grow in tetracycline medium probably contain the Drosophila DNA insert.

(c)

Expert Solution

Summary Introduction

To determine: The reason that some colonies can grow on tetracycline medium as well as ampicillin medium.

Introduction: Plasmids have an origin of replication (ori) site, which makes it possible to produce several hundred copies of a plasmid in a single host cell.

Explanation of Solution

When performing cloning process, it is not necessary for incorporation of all DNA plasmids need to be cloned. A plasmid cut with a restriction enzyme generating sticky ends can self-ligate if cut ends of the plasmid rejoin. Also, if cleavage with PstI was incomplete, then no change in biological characteristics of the uncut plasmid would be expected.

Thus, self-ligation of plasmid and incomplete cleavage with PstI are responsible for the growth of colonies that are resistant to both antibiotics.

Want to see more full solutions like this?

Subscribe now to access step-by-step solutions to millions of textbook problems written by subject matter experts!

Answer 4

Textbook Question

Chapter 20, Problem 1NST

A plasmid that is both ampicillin and tetracycline resistant is cleaved with Pstl, which cleaves within the ampicillin resistance gene. The cut plasmid is ligated with Pstl-digested Drosophila DNA to prepare a genomic library, and the mixture is used to transform E. coli K12.

Chapter 20, Problem 1NST, A plasmid that is both ampicillin and tetracycline resistant is cleaved with Pstl, which cleaves

(a) Which antibiotic should be added to the medium to select cells that have incorporated a plasmid?
(b) If recombinant cells were plated on medium containing ampicillin or tetracycline and medium with both antibiotics, on which plates would you expect to see growth of bacteria containing plasmids with Drosophila DNA inserts?
(c) How can you explain the presence of colonies that are resistant to both antibiotics?

(a)

Expert Solution

Summary Introduction

To determine: The antibiotic that an individual should add to the medium to select cells that contain the recombinant plasmid.

Introduction: Plasmid refers to an circular, extrachromosomal DNA (deoxyribonucleic acid) molecule that replicates chromosome independently.

Explanation of Solution

The gene that confers resistance to tetracycline is intact in the recombinant plasmid. An individual would add tetracycline to the medium to select the recombinant plasmd in the cells. In the medium, the bacteria that have been transformed with recombinant plasmid will be resistant to tetracycline.

Thus, an individual would use tetracycline to select cells that contain the recombinant plasmid.

(b)

Expert Solution

Summary Introduction

To determine: The plates on which an individual might see the growth of bacteria having plasmids with Drosophila DNA inserts.

Introduction: Genetically modified bacterial plasmids were the first developed vectors, used for cloning purpose.

Explanation of Solution

Cloning is a important process in which bacterial cells containing recombinant DNA can be readily identified. This process is also accomplished is through the use of selectable marker genes. Antibiotic resistant genes provide very effective selectable marker genes. Therefore, the colonies that grow on tetracycline medium but do not grow on ampicillin medium would contain the Drosophila DNA insert.

Thus, colonies that only grow in tetracycline medium probably contain the Drosophila DNA insert.

(c)

Expert Solution

Summary Introduction

To determine: The reason that some colonies can grow on tetracycline medium as well as ampicillin medium.

Introduction: Plasmids have an origin of replication (ori) site, which makes it possible to produce several hundred copies of a plasmid in a single host cell.

Explanation of Solution

When performing cloning process, it is not necessary for incorporation of all DNA plasmids need to be cloned. A plasmid cut with a restriction enzyme generating sticky ends can self-ligate if cut ends of the plasmid rejoin. Also, if cleavage with PstI was incomplete, then no change in biological characteristics of the uncut plasmid would be expected.

Thus, self-ligation of plasmid and incomplete cleavage with PstI are responsible for the growth of colonies that are resistant to both antibiotics.

Want to see more full solutions like this?

Subscribe now to access step-by-step solutions to millions of textbook problems written by subject matter experts!

Answer 5

Textbook Question

Answer 6

Textbook Question

Answer 7

(a)

Expert Solution

Summary Introduction

To determine: The antibiotic that an individual should add to the medium to select cells that contain the recombinant plasmid.

Introduction: Plasmid refers to an circular, extrachromosomal DNA (deoxyribonucleic acid) molecule that replicates chromosome independently.

Explanation of Solution

The gene that confers resistance to tetracycline is intact in the recombinant plasmid. An individual would add tetracycline to the medium to select the recombinant plasmd in the cells. In the medium, the bacteria that have been transformed with recombinant plasmid will be resistant to tetracycline.

Thus, an individual would use tetracycline to select cells that contain the recombinant plasmid.

(b)

Expert Solution

Summary Introduction

To determine: The plates on which an individual might see the growth of bacteria having plasmids with Drosophila DNA inserts.

Introduction: Genetically modified bacterial plasmids were the first developed vectors, used for cloning purpose.

Explanation of Solution

Cloning is a important process in which bacterial cells containing recombinant DNA can be readily identified. This process is also accomplished is through the use of selectable marker genes. Antibiotic resistant genes provide very effective selectable marker genes. Therefore, the colonies that grow on tetracycline medium but do not grow on ampicillin medium would contain the Drosophila DNA insert.

Thus, colonies that only grow in tetracycline medium probably contain the Drosophila DNA insert.

(c)

Expert Solution

Summary Introduction

To determine: The reason that some colonies can grow on tetracycline medium as well as ampicillin medium.

Introduction: Plasmids have an origin of replication (ori) site, which makes it possible to produce several hundred copies of a plasmid in a single host cell.

Explanation of Solution

When performing cloning process, it is not necessary for incorporation of all DNA plasmids need to be cloned. A plasmid cut with a restriction enzyme generating sticky ends can self-ligate if cut ends of the plasmid rejoin. Also, if cleavage with PstI was incomplete, then no change in biological characteristics of the uncut plasmid would be expected.

Thus, self-ligation of plasmid and incomplete cleavage with PstI are responsible for the growth of colonies that are resistant to both antibiotics.

Answer 8

(a)

Expert Solution

Summary Introduction

To determine: The antibiotic that an individual should add to the medium to select cells that contain the recombinant plasmid.

Introduction: Plasmid refers to an circular, extrachromosomal DNA (deoxyribonucleic acid) molecule that replicates chromosome independently.

Explanation of Solution

The gene that confers resistance to tetracycline is intact in the recombinant plasmid. An individual would add tetracycline to the medium to select the recombinant plasmd in the cells. In the medium, the bacteria that have been transformed with recombinant plasmid will be resistant to tetracycline.

Thus, an individual would use tetracycline to select cells that contain the recombinant plasmid.

Answer 9

(a)

Expert Solution

Answer 10

(a)

Expert Solution

Answer 11

Expert Solution

Answer 12

Summary Introduction

To determine: The antibiotic that an individual should add to the medium to select cells that contain the recombinant plasmid.

Introduction: Plasmid refers to an circular, extrachromosomal DNA (deoxyribonucleic acid) molecule that replicates chromosome independently.

Answer 13

Explanation of Solution

The gene that confers resistance to tetracycline is intact in the recombinant plasmid. An individual would add tetracycline to the medium to select the recombinant plasmd in the cells. In the medium, the bacteria that have been transformed with recombinant plasmid will be resistant to tetracycline.

Thus, an individual would use tetracycline to select cells that contain the recombinant plasmid.

Answer 14

(b)

Expert Solution

Summary Introduction

To determine: The plates on which an individual might see the growth of bacteria having plasmids with Drosophila DNA inserts.

Introduction: Genetically modified bacterial plasmids were the first developed vectors, used for cloning purpose.

Explanation of Solution

Cloning is a important process in which bacterial cells containing recombinant DNA can be readily identified. This process is also accomplished is through the use of selectable marker genes. Antibiotic resistant genes provide very effective selectable marker genes. Therefore, the colonies that grow on tetracycline medium but do not grow on ampicillin medium would contain the Drosophila DNA insert.

Thus, colonies that only grow in tetracycline medium probably contain the Drosophila DNA insert.

Answer 15

(b)

Expert Solution

Answer 16

(b)

Expert Solution

Answer 17

Expert Solution

Answer 18

Summary Introduction

To determine: The plates on which an individual might see the growth of bacteria having plasmids with Drosophila DNA inserts.

Introduction: Genetically modified bacterial plasmids were the first developed vectors, used for cloning purpose.

Answer 19

Explanation of Solution

Cloning is a important process in which bacterial cells containing recombinant DNA can be readily identified. This process is also accomplished is through the use of selectable marker genes. Antibiotic resistant genes provide very effective selectable marker genes. Therefore, the colonies that grow on tetracycline medium but do not grow on ampicillin medium would contain the Drosophila DNA insert.

Thus, colonies that only grow in tetracycline medium probably contain the Drosophila DNA insert.

Answer 20

(c)

Expert Solution

Summary Introduction

To determine: The reason that some colonies can grow on tetracycline medium as well as ampicillin medium.

Introduction: Plasmids have an origin of replication (ori) site, which makes it possible to produce several hundred copies of a plasmid in a single host cell.

Explanation of Solution

When performing cloning process, it is not necessary for incorporation of all DNA plasmids need to be cloned. A plasmid cut with a restriction enzyme generating sticky ends can self-ligate if cut ends of the plasmid rejoin. Also, if cleavage with PstI was incomplete, then no change in biological characteristics of the uncut plasmid would be expected.

Thus, self-ligation of plasmid and incomplete cleavage with PstI are responsible for the growth of colonies that are resistant to both antibiotics.

Answer 21

(c)

Expert Solution

Answer 22

(c)

Expert Solution

Answer 23

Expert Solution

Answer 24

Summary Introduction

To determine: The reason that some colonies can grow on tetracycline medium as well as ampicillin medium.

Introduction: Plasmids have an origin of replication (ori) site, which makes it possible to produce several hundred copies of a plasmid in a single host cell.

Answer 25

Explanation of Solution

When performing cloning process, it is not necessary for incorporation of all DNA plasmids need to be cloned. A plasmid cut with a restriction enzyme generating sticky ends can self-ligate if cut ends of the plasmid rejoin. Also, if cleavage with PstI was incomplete, then no change in biological characteristics of the uncut plasmid would be expected.

Thus, self-ligation of plasmid and incomplete cleavage with PstI are responsible for the growth of colonies that are resistant to both antibiotics.