PRINCIPLES OF GENE MANIPULATION (PGM), BIOL417
Fall, 2005
Dr. S. Sharp
Phone: (323) 343-2072
email: ssharp@calstatela.edu
Office: 363 BioS
Hrs:
M
10:40-12:20
T
4-5
Prerequisites: General Genetics or Microbial Genetics AND Cell Biology or Biochemistry (or their
equivalents from another university)
Final Examination: Wednesday, December 7, 8 (not 9) -10:30 AM
PGM WebPage:
link from http://www.calstatela.edu/faculty/ssharp
Required Textbooks:
T.A. Brown, Gene Cloning, An introduction. FOURTH edition. Chapman and Hall,
NY, 1995 and reprinted by Stanley Thornes (Publishers) Ltd., Cheltenham, UK,1998.
Course Reader for Biology 417, 2005
Additional items from reading list on reserve.
Helpful, on reserve:
Watson, Gilman, Witkowski, and Zoller, Recombinant DNA. Second edition.
Scientific American Books, New York, 1992.
Sample Problems
Required materials:
Two bluebooks for quizzes
GOALS FOR THE COURSE
1.
2.
3.
To gain an understanding of the concepts of gene manipulation and its applications by achieving
the objectives for each of the ten weeks.
To become familiar with the use of some of the database search tools of bioinformatics.
To introduce yourselves and each other to the breadth of career opportunities in biotechnology via
student PowerPoint presentations, presented as posters.
READING LIST / TIMELINE OF COURSE
(Items marked with an asterisk (*) are not in your course reader. Unmarked items are in your course
reader. All listed items are available at the reserve desk in the library. One of the asterisked items is
available on line at the URL cited. Note: Textbook assignments are found in your weekly lists of
objectives. They are not listed below.)
Friday, September 23: Introduction
Week 1.
Week 2.
What’s gene manipulation and why should I care? Restriction endonuclease basics.
Self-review.
1.
Pennisi, Elizabeth, “So many choices, so little money,” Science (2001), v. 294, 82-85.
2.
Butler, Declan, “Are you ready for the revolution?” Nature (2001), v. 409, 758-760.
How do you construct a cDNA clone? Gel electrophoresis basics.
Week 3.
Gel blot hybridization. Labeling, hybridization, visualization.
Week 4.
What are recombinant DNA libraries? Colony/plaque hybridization. Subcloning.
Refinements of blot hybridization analysis.
Week 5.
A wide range of vectors for your cloning and subcloning pleasure!
Midterm
Week 6.
Sequencing. PCR, DNA fingerprinting. DNA footprinting. Reporter assays.
Week 7.
Expression systems for research and profit
Week 8.
3.
Schmidt, F.R., “Recombinant expression systems in the pharmaceutical industry,” Applied
Microbiology and Biotechnology (2004) 65:363-372.
4.
Betton, J.-M., “High throughput cloning and expression strategies for protein production,”
Biochimie (2004) 86:601-605.
Stable gene transfer into animals (and plants). What’s a knock-in? a knock-out?
5.
Week 9.
Week 10.
Capecchi, Mario R., "Altering the Genome by Homologous Recombination," Science (1989),
v. 244, 1288-1292.
How do you find a gene about which you know almost nothing? Disease genes and
gene therapy.
6.*
White, R.and J.-M. Lalouel, "Chromosome Mapping with DNA Markers." Scientific
American, February, 1988, pp. 40-48.
7.
Marx, J. L., "The Cystic Fibrosis Gene is Found." Science (1989)245: 923-25.
8.
Anderson, W.F., “Human gene therapy.” (1998) Nature 392 Supplement:25-30
The post-genomic era and functional genomics
9.
Baltimore, D., “Our genome unveiled.” (2001) Nature 409:814-816.
10.
Rubin, G.M., “Comparing species.” (2001) Nature 409:820-821.
11.
Chakravarti, A., “. . . to a future of genetic medicine.” (2001) Nature 409:822-823.
Resource for term assignment
12.*
Careers in Biotechnology
Special reading assignment for final exam
13.*
Biotechnology for the 21st Century, a Report from the Biotechnology Research
Subcommittee, National Science and Technology Council. Available on the WWW at
http://www.nalusda.gov/bic/bio21
Remember to use the Table of Contents and Index to find material in your text or in the additional text on reserve to
look for unassigned material that may add to your understanding.
Please do us all a favor and bring to class or office hours any relevant current reading material you find!