Textbook Question
Compare methods for constructing homologous recombinant transgenic mice and yeast.
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18. Molecular Genetic Tools
Genetic Cloning
Back18. Molecular Genetic Tools
Genetic Cloning
- Textbook Question
What are the advantages of using a restriction enzyme whose recognition site is relatively rare? When would you use such enzymes?
807views - Textbook QuestionChimeric gene-fusion products can be used for medical or industrial purposes. One idea is to produce biological therapeutics for human medical use in animals from which the products can be easily harvested—in the milk of sheep or cattle, for example. Outline how you would produce human insulin in the milk of sheep.462views
- Textbook Question
Outline the roles played by restriction enzymes and vectors in cloning DNA.
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In 1975, the Asilomar Conference on Recombinant DNA was organized by Paul Berg, a pioneer of recombinant DNA technology, at a conference center at Asilomar State Beach in California. Physicians, scientists, lawyers, ethicists, and others gathered to draft guidelines for safe applications of recombinant DNA technology. These general guidelines were adopted by the federal government and are still in practice today. Consider the implications of recombinant DNA as a new technology. What concerns might the scientific community have had then about recombinant DNA technology? Might those same concerns exist today?
914views - Textbook QuestionWhy are diseases of the blood simpler targets for treatment by gene therapy than are many other genetic diseases?419views
- Textbook Question
Injection of double-stranded RNA can lead to gene silencing by degradation of RNA molecules complementary to either strand of the dsRNA. Could RNAi be used in gene therapy for a defect caused by a recessive allele? A dominant allele? If so, what might be the major obstacle to using RNAi as a therapeutic agent?
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What are some of the impacts of biotechnology on crop plants in the United States?
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In the context of recombinant DNA technology, of what use is a probe?
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Summarize the arguments for and against patenting genetically modified organisms.
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If you performed a PCR experiment starting with only one copy of double-stranded DNA, approximately how many DNA molecules would be present in the reaction tube after 15 cycles of amplification?
714views - Textbook QuestionIt is often desirable to insert cDNAs into a cloning vector in such a way that all the cDNA clones will have the same orientation with respect to the sequences of the plasmid. This is referred to as directional cloning. Outline how you would directionally clone a cDNA library in the plasmid vector pUC18.678views
- Textbook QuestionA major advance in the 1980s was the development of technology to synthesize short oligonucleotides. This work both facilitated DNA sequencing and led to the advent of the development of PCR. Recently, rapid advances have occurred in the technology to chemically synthesize DNA, and sequences up to 10 kb are now readily produced. As this process becomes more economical, how will it affect the gene-cloning approaches outlined in this chapter? In other words, what types of techniques does this new technology have potential to supplant, and what techniques will not be affected by it?695views
- Textbook Question
The bacteriophage lambda genome can exist in either a linear form or a circular form.
How many fragments will be formed by restriction enzyme digestion with XhoI alone, with XbaI alone, and with both XhoI and XbaI in the linear and circular forms of the lambda genome?
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You have recovered a cloned DNA segment from a vector and determine that the insert is 1300 bp in length. To characterize this cloned segment, you isolate the insert and decide to construct a restriction map. Using enzyme I and enzyme II, followed by gel electrophoresis, you determine the number and size of the fragments produced by enzymes I and II alone and in combination, as recorded in the following table. Construct a restriction map from these data, showing the positions of the restriction-enzyme cutting sites relative to one another and the distance between them in units of base pairs.
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