18. Molecular Genetic Tools

Genetic Cloning

Open Question
Using animal models of human diseases can lead to insights into the cellular and genetic bases of the diseases. Duchenne muscular dystrophy (DMD) is the consequence of an X-linked recessive allele.How would you make a Drosophila model of DMD?
Open 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?
Open Question
Using animal models of human diseases can lead to insights into the cellular and genetic bases of the diseases. Duchenne muscular dystrophy (DMD) is the consequence of an X-linked recessive allele.How would you make a mouse model of DMD?
Open Question
List the advantages and disadvantages of using plasmids as cloning vectors. What advantages do BACs and YACs provide over plasmids as cloning vectors?
Open Question
Compare methods for constructing homologous recombinant transgenic mice and yeast.
Open Question
What are the advantages of using a restriction enzyme whose recognition site is relatively rare? When would you use such enzymes?
Open Question
Chimeric 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.
Open Question
Outline the roles played by restriction enzymes and vectors in cloning DNA.
Open Question
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?
Open Question
Why are diseases of the blood simpler targets for treatment by gene therapy than are many other genetic diseases?
Open 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 (see Sections 13.3 and 14.3) 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?
Open Question
What are some of the impacts of biotechnology on crop plants in the United States?
Open Question
In the context of recombinant DNA technology, of what use is a probe?
Open Question
Summarize the arguments for and against patenting genetically modified organisms.
Open Question
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?