COURSE TITLE Biology 303; Microbial Genetics
PROFESSOR(S) Jeff Knight, jknight@mtholyoke.edu
LECTURE  
LAB 3 Hour Lab Required
TEXTS

Griffiths, AJF, Lewontin, RC, Gelbart, WM. 2002. Modern Genetic Analysis. 2nd ed. W. H. Freeman Company.

EXAMS & GRADING

Perhaps no area of biology has changed so dramatically in the last twenty years or so as has the field of molecular genetics. While it is true that molecular biology today involves the study of nucleic acids, proteins, and organelles from all forms of life, it is also true that most of the groundwork for this study was laid by extensive work on the molecular genetics of bacteria and their viruses. Prokaryotic systems continue to provide exciting experimental challenges to investigators throughout the world today.

Molecular genetics is taught in four upper level courses at Mount Holyoke: this one, Mr. Woodard's eukaryotic molecular genetics class (Biology 340), and Biochemistry 314 and 330. The material in this course will concentrate on the molecular genetics of microbial systems (bacteria, viruses, and simple eukaryotic systems such as yeast), while other eukaryotic systems will be stressed in Biology 340 and Biochemistry 314 and 330.

It is only natural that one of your first concerns will be the manner in which your course grade will be determined. Here's the story:

Requirement
% of Grade
Midterm Exams (3)
40
Laboratory Work
20
Research Proposal
20
Oral Defense
10
Class Discussions
10
Total
100

The first exam will cover approximately the first third of the lecture material, and it will be given in class on Wednesday, February 26. The second will be given in class on Wednesday, April 2, and it will cover material presented since the first exam. The third will be given on Wednesday, April 30. There will be no comprehensive final examination.

The fun part of the course will be your development of an original research proposal that you have designed to answer a particular question that you have determined to be of interest to you concerning some specific experimental system in the field. The written proposal will be due on our last day of classes, Monday, May 5. You will then have the opportunity to orally defend your proposal against any questions or criticisms I might have. We will schedule 30 minute meetings for this purpose, at your individual convenience, during the reading and final exam periods.

On most Fridays, we will spend our time together discussing papers from the primary literature. Twice during the semester, each student will have the responsibility of presenting the week's paper and leading the discussion. I expect these to be informed and critical discussions, with presenters serving as the knowledgeable authorities. Evaluation for this part of the course will depend both on the quality of your two presentations and your input to discussions when other students are presenting.

LECTURE SCHEDULE
Major Topic
Textbook Reading
I. Introduction to the Molecular Genetics of Microbial Systems
A. The prokaryotic cell  
B. The bacteriophage pp. 283 - 285
C. Biochemical Methodology
    1. Isolation of DNA
    2. CsCl and sucrose gradients
    3. Density labeling
    4. DNA hybridization
    5. Electrophoresis
    6. Restriction endonuclease analysis
    7. Southern, Western, and Northern blot analysis
    8. The Polymerase Chain Reaction and DNA sequencing
    9. Recombinant DNA technology
        a) choice of vector
        b) new applications and variations
Chapter 10
II. Genetic Manipulations in Microbial Systems
A. Bacterial transformation
B. Bacterial conjugation (episome transfer)
C. F-duction
D. Transduction
Chapter 9
III. Properties of DNA and RNA
A. DNA replication
    1. Enzymes and proteins involved
    2. Models
    3. Experimental evidence
    4. Reverse transcriptase
pp. 86 - 91
B. The nature of mutation and mutagenesis
    1. Site-directed mutagenesis
    2. Mutagenic PCR
Chapter 7
pp. 342 - 343
C. Genetic fine structure
D. Host restriction and modification
E. DNA repair
F. Recombination
G. The genetic code and its ramifications
    1. Nature of the code
    2. RNA polymerase and transcription
    3. Errors in protein synthesis - suppression







pp. 177 - 178
H. Transposable elements in bacteria Chapter 13
IV. Regulation in Prokaryotic Systems
A. The operon
    1. Inducible systems
    2. Repressible systems
B. The regulon
C. Positive control systems
pp. 433 - 448
LAB SCHEDULE
I expect that each of you will maintain a laboratory notebook in which you will record procedures, data, and conclusions while you are performing the experiments. People's styles with regards to documenting laboratory work in a notebook differ greatly, and I won't presume to impose mine on you. I will, however, be observing (and taking notes) on the manner in which you generate, record, and interpret your data in your notebook. In addition, you are expected to turn in three laboratory reports during the semester. These can be about any of the laboratory exercises you do except experiments 2 and 8. The reports are due two weeks after the laboratory work is completed or by May 6, whichever comes first. College policy dictates that all written work should be turned in by the last day of classes. Hopefully, many of you will choose to get your laboratory reports done and out of the way early in the semester. I strongly urge that you study the purpose and procedures of each experiment before you come to the laboratory. This saves time and confusion and often prevents the occurrence of silly mistakes.

Week
Topic
1 Experiment 1. Bacterial Growth
2 Experiment 2. Preparation of Phage T4
3

Experiment 3. Complementation and Recombination in Phage T4

4 Experiment 4. Stock Construction
5 Experiment 5. Biochemical Characterization of Auxotrophic Mutants
6 Experiment 6. UV-Induced Mutations in E. coli
7 NO LAB
8 Experiment 7. Rec- Strains
Experiment 8. Episome Curing
9 Experiment 9. Genetic and Biochemical Characterization of Lac- Mutants in E. coli
10 Experiment 9 continued.
Experiment 10. Yeast Genetics I
11 Experiment 10. Yeast Genetics II
12 Experiment 10. Yeast Genetics III
13 Experiment 10. Yeast Genetics IV
14 NO LAB

Last Modified: March 11, 2005

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