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Ch. 20 - Recombinant DNA Technology
All textbooksKlug 12th EditionCh. 20 - Recombinant DNA TechnologyProblem 5
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Chapter 19, Problem 5

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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Key Concepts

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

Recombinant DNA Technology

Recombinant DNA technology involves combining DNA from different sources to create new genetic combinations. This technique allows scientists to manipulate genes and produce proteins, which can be used in various applications, including medicine and agriculture. Understanding this technology is crucial for recognizing why different host cells are utilized in cloning.
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Recombination after Single Strand Breaks

Host Cell Selection

The choice of host cell for recombinant DNA is influenced by factors such as the desired protein's complexity, post-translational modifications, and growth conditions. While bacteria are commonly used due to their rapid growth and simplicity, eukaryotic cells like yeast or mammalian cells may be preferred for producing proteins that require specific modifications or are toxic to bacteria.
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Cell-cell interactions

Applications of Cloning

Cloning has diverse applications beyond bacterial systems, including gene therapy, vaccine production, and the creation of genetically modified organisms (GMOs). Different cell types can provide unique advantages, such as higher yields of complex proteins or the ability to study gene function in a more relevant biological context, making the use of various hosts essential in biotechnology.
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Textbook Question

The human insulin gene contains a number of sequences that are removed in the processing of the mRNA transcript. In spite of the fact that bacterial cells cannot excise these sequences from mRNA transcripts, explain how a gene like this can be cloned into a bacterial cell and produce insulin.

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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. 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

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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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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