Lecture 30: PCR, knockout and transgenic organisms, biotechnology
1. The polymerase chain reaction (PCR) is a widely used technique in
molecular genetics.
2. One of the ways of determining whether a sequence has been
successfully amplified by PCR is to do electrophoresis and look
for a band of DNA of sharply defined size. Explain how such a
band arises during PCR.
3. Why is it not necessary to add new DNA polymerase and primers
for each new cycle of PCR?
4. You have a cDNA of unknown sequence in a plasmid. How could
you do a PCR amplification of the cDNA without first determining
its end sequences?
5. What types of precautions must be taken when doing PCR to be
certain that the desired results are obtained?
6. What aspects of PCR make it particularly useful in forensic
investigations? How do the same properties make PCR particularly
susceptible to challenge by defense lawyers?
7. What are the essential features that must be included in a
yeast artificial chromosome (YAC)?
8. What types of vectors are particularly useful for transformation
of animal cells?
9. What types of vectors are particularly useful for transformation
of plants?
10. How are transgenic mice generated?
11. Describe the various techniques that can be used to introduce
a foreign DNA into a cell.
12. What is a knockout mouse and what is its special research
value?
13. What is a reporter gene and what is it used for? Briefly describe
threee commonly used reporter genes.
14. What type of genetic modification has been made in the Flavr
Savr tomato? What is the market value of such a change?
15. What types of genetic diseases should be the most easily treated
by genetic engineering?
16. What characteristics must a protein have to make it a practical
candidate for commercial manufacture using recombinant DNA technology?
17. What are the practical reasons that often make it desirable
to use an inducible promoter for commercial production of proteins
from genetically engineered cells?
18. What problems make it impractical to produce some types of
proteins using prokaryotic expression vector systems?
19. Cite an example of the spread of a human infectious disease
by proteins derived from human sources that has been eliminated
through the use of biotechnology. Identify both the protein and
the disease.
20. Cite an example of a human genetic disease that has been successfully
treated with gene therapy.
21. A substantial effort is being devoted to making certain kinds
of plants resistant to herbicides such as glycophosphate.
22. Describe the use of genetically engineered dairy animals for
the production of non-milk proteins. What advantages does this
procedure offer over alternative forms of biotechnology?
23. You have cloned a cDNA in a pBR322 vector. You now want to
produce the protein coded by that cDNA in large amounts. What
additional steps would be necessary?
24. What is an embryonal stem cell (ES cell) and what is its special
value in genetic research?
25. Describe two different approaches that are being taking toward
the use of biotechnology in vaccine production.
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a. What special property must be possessed by the DNA polymerase
used in PCR, and why is this so important for doing PCR?
b. Explain the use of primers in PCR, including why two different primers
must be used.
c. What relationship must exist between the two primers used in PCR?
d. Summarize all of the components that must be present in a PCR
reaction mixture.
e. Summarize the overall process that makes possible extensive amplification
of specific DNA sequences with minimal effort through the use of PCR.
Questions below this line will not be included on the fourth examination
on November 18, 1998.
a. What is the economic advantage of such a plant?
b. What procedures are being used?
c. What potential environmental concerns have been expressed about
widespread use of such plants?
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