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Ch. 20 - Recombinant DNA Technology
Chapter 19, Problem 6

Using DNA sequencing on a cloned DNA segment, you recover the nucleotide sequence shown below. Does this segment contain a palindromic recognition sequence for a restriction enzyme? If so, what is the double-stranded sequence of the palindrome, and what enzyme would cut at this sequence? (Consult Figure 20.1 for a list of restriction sites.)

CAGTATGGATCCCAT

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1
Identify the given DNA sequence: CAGTATGGATCCCAT.
Understand that a palindromic sequence in DNA is a sequence that reads the same in the 5' to 3' direction on both strands.
Write the complementary strand of the given sequence: GTCATACCTAGGGTA.
Check for palindromic sequences by comparing the sequence and its complement for symmetry.
Consult a list of known restriction enzyme recognition sites to match any identified palindromic sequence with a specific enzyme.

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Key Concepts

Here are the essential concepts you must grasp in order to answer the question correctly.

Palindromic Sequences

A palindromic sequence in DNA is a sequence of nucleotides that reads the same in both directions on complementary strands. This characteristic is crucial for the recognition of specific sites by restriction enzymes, which often require such symmetry to bind and cut the DNA effectively.
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Restriction Enzymes

Restriction enzymes, or restriction endonucleases, are proteins that cut DNA at specific sequences, typically palindromic sites. Each enzyme recognizes a unique sequence, allowing for targeted manipulation of DNA, which is essential in molecular biology for cloning and genetic engineering.
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DNA Sequencing

DNA sequencing is the process of determining the precise order of nucleotides within a DNA molecule. In this context, sequencing the cloned DNA segment allows for the identification of specific sequences, including potential palindromic sites that may be recognized by restriction enzymes.
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Related Practice
Textbook Question
One of the major causes of sickness, death, and economic loss in the cattle industry is Mannheimia haemolytica, which causes bovine pasteurellosis, or shipping fever. Noninvasive delivery of a vaccine using transgenic plants expressing immunogens would reduce labor costs and trauma to livestock. An early step toward developing an edible vaccine is to determine whether an injected version of an antigen (usually a derivative of the pathogen) is capable of stimulating the development of antibodies in a test organism. The following table assesses the ability of a transgenic portion of a toxin (Lkt) of M. haemolytica to stimulate development of specific antibodies in rabbits. Immunogen Injected Antibody Production in Serum Lkt50*—saline extract + Lkt50*—column extract + Mock injection - *Lkt50 is a smaller derivative of Lkt that lacks all hydrophobic regions. indicates at least 50 percent neutralization of toxicity of Lkt; indicates no neutralization activity. Source: Modified from Lee et al. (2001). Infect. and Immunity 69:5786–5793. What general conclusion can you draw from the data?
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Textbook Question
One of the major causes of sickness, death, and economic loss in the cattle industry is Mannheimia haemolytica, which causes bovine pasteurellosis, or shipping fever. Noninvasive delivery of a vaccine using transgenic plants expressing immunogens would reduce labor costs and trauma to livestock. An early step toward developing an edible vaccine is to determine whether an injected version of an antigen (usually a derivative of the pathogen) is capable of stimulating the development of antibodies in a test organism. The following table assesses the ability of a transgenic portion of a toxin (Lkt) of M. haemolytica to stimulate development of specific antibodies in rabbits. Immunogen Injected Antibody Production in Serum Lkt50*—saline extract + Lkt50*—column extract + Mock injection - *Lkt50 is a smaller derivative of Lkt that lacks all hydrophobic regions. indicates at least 50 percent neutralization of toxicity of Lkt; indicates no neutralization activity. Source: Modified from Lee et al. (2001). Infect. and Immunity 69:5786–5793. With regards to development of a usable edible vaccine, what work remains to be done?
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Textbook Question

Although many cloning applications involve introducing recombinant DNA into bacterial host cells, many other cell types are also used as hosts for recombinant DNA. Why?

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Textbook Question

As genetic testing becomes widespread, medical records will contain the results of such testing. Who should have access to this information? Should employers, potential employers, or insurance companies be allowed to have this information? Would you favor or oppose having the government establish and maintain a central database containing the results of individuals' genome scans?

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Textbook Question

Restriction sites are palindromic; that is, they read the same in the 5' to 3' direction on each strand of DNA. What is the advantage of having restriction sites organized this way?

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Textbook Question

Might it make sense someday to sequence every newborn's genome at the time of birth? What are the potential advantages and concerns of this approach?

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