Chapter 10, Problem 2Q

(A)

Summary Introduction

To explain: Digestion products by the enzyme EcoRI.

Introduction:

Enzymes that recognize and cleave specific sites of the DNA molecule are called restriction enzymes. When the cleavage occurs at the end of the DNA molecule, it is referred to as exonucleases and when the cleavage occurs within the DNA molecule, it is called endonucleases. Restriction enzymes were first discovered in bacteria. A bacterium contains restriction enzymes within it to degrade the foreign DNA entering the cell and protects itself from enzymes by chemically modifying its own DNA.

Different bacteria contain different enzymes that will have unique cleavage properties that only cleave the DNA at specific sites. In recombinant DNA technology, restriction enzymes are isolated from different bacteria and are utilized to cut the desired DNA at specific sites. The enzymes cleave the sequences differently. The enzyme cleaves the sequences either by sticky ends or by blunt ends. Blunt ends are formed when the enzymes make a straight cut in the given sequences. The sticky ends are characterized by a cut made by the enzyme producing a staggered end. Different enzymes cleave the given sequence variably. EcoRI and Hind III produce staggered (sticky ends) ends while Hae III makes blunt ends.

Given sequence:

5'-AAGAATTGCGGAATTCGGGCCTTAAGCGCCGCGTCGAGGCCTTAAA- 3'

3' –TTCTTAACGCCTTAAGCCCGGAATTCGCGGCGCAGCTCCGGAATTT- 5'

(B)

Summary Introduction

To explain: Digestion products by the enzyme Hae III

Introduction:

Enzymes that recognize and cleave specific sites of the DNA molecule are called restriction enzymes. When the cleavage occurs at the end of the DNA molecule, it is referred to as exonucleases and when the cleavage occurs within the DNA molecule, it is called endonucleases. Restriction enzymes were first discovered in bacteria. A bacterium contains restriction enzymes within it to degrade the foreign DNA entering the cell and protects itself from enzymes by chemically modifying its own DNA.

Different bacteria contain different enzymes that will have unique cleavage properties that only cleave the DNA at specific sites. In recombinant DNA technology, restriction enzymes are isolated from different bacteria and are utilized to cut the desired DNA at specific sites. The enzymes cleave the sequences differently. The enzyme cleaves the sequences either by sticky ends or by blunt ends. Blunt ends are formed when the enzymes make a straight cut in the given sequences. The sticky ends are characterized by a cut made by the enzyme producing a staggered end. Different enzymes cleave the given sequence variably. EcoRI and Hind III produce staggered (sticky ends) ends while Hae III makes blunt ends.

Given sequence:

5'-AAGAATTGCGGAATTCGGGCCTTAAGCGCCGCGTCGAGGCCTTAAA- 3'

3' –TTCTTAACGCCTTAAGCCCGGAATTCGCGGCGCAGCTCCGGAATTT- 5'

(C)

Summary Introduction

To explain: Digestion products by the enzyme Hind III

Introduction:

Enzymes that recognize and cleave specific sites of the DNA molecule are called restriction enzymes. When the cleavage occurs at the end of the DNA molecule, it is referred to as exonucleases and when the cleavage occurs within the DNA molecule, it is called endonucleases. Restriction enzymes were first discovered in bacteria. A bacterium contains restriction enzymes within it to degrade the foreign DNA entering the cell and protects itself from enzymes by chemically modifying its own DNA.

Different bacteria contain different enzymes that will have unique cleavage properties that only cleave the DNA at specific sites. In recombinant DNA technology, restriction enzymes are isolated from different bacteria and are utilized to cut the desired DNA at specific sites. The enzymes cleave the sequences differently. The enzyme cleaves the sequences either by sticky ends or by blunt ends. Blunt ends are formed when the enzymes make a straight cut in the given sequences. The sticky ends are characterized by a cut made by the enzyme producing a staggered end. Different enzymes cleave the given sequence variably. EcoRI and Hind III produce staggered (sticky ends) ends while Hae III makes blunt ends.

Given sequence:

5'-AAGAATTGCGGAATTCGGGCCTTAAGCGCCGCGTCGAGGCCTTAAA- 3'

3' –TTCTTAACGCCTTAAGCCCGGAATTCGCGGCGCAGCTCCGGAATTT- 5'

(D)

Summary Introduction

To explain: Digestion product produced when all the three enzymes are combined.

Introduction:

Enzymes that recognize and cleave specific sites of the DNA molecule are called restriction enzymes. When the cleavage occurs at the end of the DNA molecule, it is referred to as exonucleases and when the cleavage occurs within the DNA molecule, it is called endonucleases. Restriction enzymes were first discovered in bacteria. A bacterium contains restriction enzymes within it to degrade the foreign DNA entering the cell and protects itself from enzymes by chemically modifying its own DNA.

Different bacteria contain different enzymes that will have unique cleavage properties that only cleave the DNA at specific sites. In recombinant DNA technology, restriction enzymes are isolated from different bacteria and are utilized to cut the desired DNA at specific sites. The enzymes cleave the sequences differently. The enzyme cleaves the sequences either by sticky ends or by blunt ends. Blunt ends are formed when the enzymes make a straight cut in the given sequences. The sticky ends are characterized by a cut made by the enzyme producing a staggered end. Different enzymes cleave the given sequence variably. EcoRI and Hind III produce staggered (sticky ends) ends while Hae III makes blunt ends.

Given sequence:

5'-AAGAATTGCGGAATTCGGGCCTTAAGCGCCGCGTCGAGGCCTTAAA- 3'

3' –TTCTTAACGCCTTAAGCCCGGAATTCGCGGCGCAGCTCCGGAATTT- 5'