(a) Interpretation: The reason by which cell C can grow in the HAT medium is to be determined. Concept introduction: HAT medium is the selection medium, used for mammalian cell culture. It is the combination of hypoxanthine, aminopterin, and thymidine. Hypoxanthine and thymidine are the intermediates formed in the DNA synthesis. While aminopterin is the inhibitor of dihydrofolate reductase.

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Answer 1

Question

Chapter 25, Problem 19P

Interpretation Introduction

(a)

Interpretation:

The reason by which cell C can grow in the HAT medium is to be determined.

Concept introduction:

HAT medium is the selection medium, used for mammalian cell culture. It is the combination of hypoxanthine, aminopterin, and thymidine. Hypoxanthine and thymidine are the intermediates formed in the DNA synthesis. While aminopterin is the inhibitor of dihydrofolate reductase.

Interpretation Introduction

(b)

Interpretation:

Distinction between the normal cells A and the cells that have taken up the foreign DNA is to be determined.

Concept introduction:

HAT medium is the selection medium, used for mammalian cell culture. It is the combination of hypoxanthine, aminopterin, and thymidine. Hypoxanthine and thymidine are the intermediates formed in the DNA synthesis. While aminopterin is the inhibitor of dihydrofolate reductase.

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The beta-lactamase hydrolyzes the lactam-ring in penicillin. Describe the mechanism of hydrolysis, insuring to include the involvement of S, D, & K in the reaction sequence. Please help

To map the active site of beta-lactamase, the enzyme was hydrolyzed with trypsin to yield a hexapeptide (P1) with the following amino acids. Glu, Lys, Leu, Phe, Met, and Ser. Treatment of P1 with phenyl isothiocyanate yielded a PTH derivative of phenylalanine and a peptide (P2). Treatment of P1 with cyanogenbromide gave an acidic tetrapeptide (P3) and a dipeptide (P4).Treatment of P2 with 1-fluoro-2,4-dinitrobenzene, followed by complete hydrolysis, yields N-2,4-dinitrophenyl-Glu. P1, P2, and P3 contain the active site serine. Why doesn't D in this hexapeptide not participate in the hydrolysis of the beta-lactam ring even though S, K, and D are involved in the catalyst?

To map the active site of -lactamase, the enzyme was hydrolyzed with trypsin to yield a hexapeptide (P1) with the following amino acids. Glu, Lys, Leu, Phe, Met, and Ser. Treatment of P1 with phenyl isothiocyanate yielded a PTH derivative of phenylalanine and a peptide (P2). Treatment of P1 with cyanogenbromide gave an acidic tetrapeptide (P3) and a dipeptide (P4).Treatment of P2 with 1-fluoro-2,4-dinitrobenzene, followed by complete hydrolysis, yields N-2,4-dinitrophenyl-Glu. P1, P2, and P3 contain the active site serine. Using the experimental results described above derive the primary sequence of the active site hexapeptide. Please help!

Answer 2

Question

Chapter 25, Problem 19P

Interpretation Introduction

(a)

Interpretation:

The reason by which cell C can grow in the HAT medium is to be determined.

Concept introduction:

HAT medium is the selection medium, used for mammalian cell culture. It is the combination of hypoxanthine, aminopterin, and thymidine. Hypoxanthine and thymidine are the intermediates formed in the DNA synthesis. While aminopterin is the inhibitor of dihydrofolate reductase.

Interpretation Introduction

(b)

Interpretation:

Distinction between the normal cells A and the cells that have taken up the foreign DNA is to be determined.

Concept introduction:

HAT medium is the selection medium, used for mammalian cell culture. It is the combination of hypoxanthine, aminopterin, and thymidine. Hypoxanthine and thymidine are the intermediates formed in the DNA synthesis. While aminopterin is the inhibitor of dihydrofolate reductase.

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Answer 3

Question

Chapter 25, Problem 19P

Interpretation Introduction

(a)

Interpretation:

The reason by which cell C can grow in the HAT medium is to be determined.

Concept introduction:

HAT medium is the selection medium, used for mammalian cell culture. It is the combination of hypoxanthine, aminopterin, and thymidine. Hypoxanthine and thymidine are the intermediates formed in the DNA synthesis. While aminopterin is the inhibitor of dihydrofolate reductase.

Interpretation Introduction

(b)

Interpretation:

Distinction between the normal cells A and the cells that have taken up the foreign DNA is to be determined.

Concept introduction:

HAT medium is the selection medium, used for mammalian cell culture. It is the combination of hypoxanthine, aminopterin, and thymidine. Hypoxanthine and thymidine are the intermediates formed in the DNA synthesis. While aminopterin is the inhibitor of dihydrofolate reductase.

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Answer 4

Question

Chapter 25, Problem 19P

Interpretation Introduction

(a)

Interpretation:

The reason by which cell C can grow in the HAT medium is to be determined.

Concept introduction:

HAT medium is the selection medium, used for mammalian cell culture. It is the combination of hypoxanthine, aminopterin, and thymidine. Hypoxanthine and thymidine are the intermediates formed in the DNA synthesis. While aminopterin is the inhibitor of dihydrofolate reductase.

Interpretation Introduction

(b)

Interpretation:

Distinction between the normal cells A and the cells that have taken up the foreign DNA is to be determined.

Concept introduction:

HAT medium is the selection medium, used for mammalian cell culture. It is the combination of hypoxanthine, aminopterin, and thymidine. Hypoxanthine and thymidine are the intermediates formed in the DNA synthesis. While aminopterin is the inhibitor of dihydrofolate reductase.

Expert Solution & Answer

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Answer 5

Chapter 25, Problem 19P

Interpretation Introduction

(a)

Interpretation:

The reason by which cell C can grow in the HAT medium is to be determined.

Concept introduction:

HAT medium is the selection medium, used for mammalian cell culture. It is the combination of hypoxanthine, aminopterin, and thymidine. Hypoxanthine and thymidine are the intermediates formed in the DNA synthesis. While aminopterin is the inhibitor of dihydrofolate reductase.

Interpretation Introduction

(b)

Interpretation:

Distinction between the normal cells A and the cells that have taken up the foreign DNA is to be determined.

Concept introduction:

HAT medium is the selection medium, used for mammalian cell culture. It is the combination of hypoxanthine, aminopterin, and thymidine. Hypoxanthine and thymidine are the intermediates formed in the DNA synthesis. While aminopterin is the inhibitor of dihydrofolate reductase.

Answer 6

Chapter 25, Problem 19P

Interpretation Introduction

(a)

Interpretation:

The reason by which cell C can grow in the HAT medium is to be determined.

Concept introduction:

HAT medium is the selection medium, used for mammalian cell culture. It is the combination of hypoxanthine, aminopterin, and thymidine. Hypoxanthine and thymidine are the intermediates formed in the DNA synthesis. While aminopterin is the inhibitor of dihydrofolate reductase.

Answer 7

Chapter 25, Problem 19P

Interpretation Introduction

(a)

Interpretation:

The reason by which cell C can grow in the HAT medium is to be determined.

Concept introduction:

HAT medium is the selection medium, used for mammalian cell culture. It is the combination of hypoxanthine, aminopterin, and thymidine. Hypoxanthine and thymidine are the intermediates formed in the DNA synthesis. While aminopterin is the inhibitor of dihydrofolate reductase.

Answer 8

Interpretation Introduction

(b)

Interpretation:

Distinction between the normal cells A and the cells that have taken up the foreign DNA is to be determined.

Concept introduction:

HAT medium is the selection medium, used for mammalian cell culture. It is the combination of hypoxanthine, aminopterin, and thymidine. Hypoxanthine and thymidine are the intermediates formed in the DNA synthesis. While aminopterin is the inhibitor of dihydrofolate reductase.

Answer 9

Interpretation Introduction

(b)

Interpretation:

Distinction between the normal cells A and the cells that have taken up the foreign DNA is to be determined.

Concept introduction:

HAT medium is the selection medium, used for mammalian cell culture. It is the combination of hypoxanthine, aminopterin, and thymidine. Hypoxanthine and thymidine are the intermediates formed in the DNA synthesis. While aminopterin is the inhibitor of dihydrofolate reductase.

Answer 10

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Answer 11

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Answer 12

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Answer 13

Ch. 25 - Prob. 1P Ch. 25 - Prob. 2P Ch. 25 - Prob. 3P Ch. 25 - Prob. 4P Ch. 25 - Prob. 5P Ch. 25 - Prob. 6P Ch. 25 - Prob. 7P Ch. 25 - Prob. 8P Ch. 25 - Prob. 9P Ch. 25 - Prob. 10P

Solution Summary: The author explains that HAT medium is the selection medium used for mammalian cell culture. It contains hypoxanthine, aminopterin, and thymidine.

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