H.
Shubeita, Ph.D. (handouts) & M.
Mardahl, Ph.D. (handouts)
Recombinant
DNA
Lecture
Schedule
Date |
Topic |
Instructor
|
Reading
assignment (O&P, 6th Ed.)
|
|
Jan.
21
|
Introduction;
Overview of Course
|
Shubeita/Mardahl
|
1-5
|
|
|
DNA
Structure and Replication
|
Shubeita
|
see
Genes VII
|
|
Jan
23
|
Molecular
Biology of Prokaryotes: a Review
|
Shubeita
|
see
Genes VII
|
|
Jan
28
|
Molecular
Biology of Eukaryotes: a Review
|
Shubeita
|
see
Genes VII
|
|
Jan
30
|
Isolation
and Purification of DNA (Genomic DNA, Plasmid DNA, Phage DNA), Agarose
and Acrylamide Gel Electrophoresis
of DNA
|
Mardahl
|
8-10,
43-44;
48-49 |
|
Feb
4
|
DNA
Restriction and Methylation Enzymes
|
Shubeita
|
|
|
Feb
6
|
Other
Enzymes used in Recombinant DNA Research
|
Shubeita
|
36-42
|
|
Feb
11
|
Restriction
Enzyme Mapping
|
Mardahl
|
277-282
|
|
Feb
13
|
Shubeita
|
||
|
Feb
18
|
Cloning
in Bacteria; Cloning Vectors: Plasmids, M13
|
Shubeita
|
|
|
Feb
20
|
FIRST
EXAM
|
Shubeita/Mardahl
|
|
|
Feb
25
|
Cloning
Vectors: Lambda, Cosmids, Phagemids
|
Shubeita
|
|
|
Feb
27
|
Techniques
for Introduction of DNA into Bacterial, Plant and Animal Cells
|
Mardahl
|
17-19;
139-143; 156-157; 176-201
|
|
Mar
4
|
Genomic
Library Construction and Screening
|
Shubeita
|
|
|
Mar
6
|
Isolation
and Analysis of RNA; Synthesis and Cloning of cDNA; cDNA libraries
|
Mardahl
|
|
|
Mar
11
|
Expression
and Purification of Recombinant Proteins
|
Shubeita
|
|
|
Mar
13
|
Use
of Computers for Analyzing Nucleic Acid and Protein Sequences, Computer
assignment, location to be announced
|
J.
Trawick, Elitra,
Shubeita |
126-132
|
|
Mar
18
|
Broad
Host Range Plasmids, Prokaryotic Gene Transfer.
|
Michael
Cleary, PC Kelco, Mardahl
|
|
|
Mar
20
|
Synthesis
and Uses of Oligonucleotides; DNA sequencing
|
Shubeita
|
|
|
Mar
25
|
PCR;
Applications of PCR
|
Shubeita
|
|
|
Mar
27
|
SECOND
EXAM
|
Shubeita/Mardahl
|
|
|
Apr
8
|
Eukaryotic
Cloning Vectors I
|
S.
Shah, Invitrogen, Shubeita
|
|
|
Apr
10
|
Eukaryotic
Cloning Vectors II
|
S.
Shah, Invitrogen, Shubeita
|
|
|
Apr
15
|
In
vitro
Mutagenesis; Reverse Genetics
|
Mardahl
|
132-138;
|
|
Apr
17
|
Defining
the Location of 5’ and 3’ Ends of Transcripts; Assays of DNA Binding Proteins
|
Shubeita
|
|
|
Apr
22
|
Gene
Therapy and Viral Vaccines
|
Mardahl
|
288-299
|
|
Apr
24
|
Transgenic
Eggs and Animals
|
Mardahl
|
201-220
|
|
Apr
29
|
Transgenic
Eggs and Animals
|
Mardahl
|
247-273
|
|
May
1
|
Studying
Eukaryotic Gene Control Using Recombinant DNA Technology
|
Shubeita
|
|
|
May
6
|
Diagnostics,
Disease Prevention/Treatment, Food Production
|
Shubeita
|
|
|
May
8
|
Human
Genome
|
Mardahl
|
126-132;
274-288
|
COURSE
REQUIREMENTS: 3
exams (100 points each), 1 written paper (100 points), 1 computer assignment
(100 points). Copies of previous exams are available on reserve at the
library. Because of the very
large amount of material covered, we will not have a comprehensive final
exam. However, the third exam
will take place during the final exam period (Tuesday, May 13, 7:00
to 9:00 pm). You will have
1.5 hours to complete each of the 1st two exams. For
the written paper, choose an article from the list of articles provided
by the instructors. The articles
utilize recombinant DNA techniques to answer basic scientific questions
or to provide a method for synthesizing an important product. Attach
one copy of the article to the term paper. You
will need to do some background reading on the subject of your article
in order to produce a satisfactory paper. Critique
the methods used in the paper based on what you have learned in this course. The
paper should be typed, double-spaced, and 5-10 pages in length. April
24 will be the last day for submission of your completed term paper. See
the following pages for more details on the term paper. The
computer exercise will be described in class on March 13 and is due April
10.
Required
Texts: Principles
of Gene Manipulation; Old and Primrose, Sixth Edition
Recent
research articles (citations will be provided in class)
Supplemental
Texts: Genes
VI or Genes VII, Lewin
Recombinant
DNA, Watson, Gilman, Witkowski, and Zoller, Second Edition
Methods
in Enzymology, Vol. 152
Review
of Journal Article
Introduction:
1. Definition
of problems and issues covered in article.
2. Discussion
of background information and previous results.
Evaluation
of Experimental Results:
1. What
are the specific questions being addressed?
2. Description
and evaluation of techniques used.
3. What
are the experimental results?
4. Evaluate
the controls that were used. Were
the authors careful to include correct controls?
5. Are
the results correctly interpreted and are the conclusions warranted ?
Discussion:
1. Evaluate
the significance of the results you described in the previous section. Place
these results in the context of the field.
2. What
are the questions that remain to be answered and what directions are most
likely to lead to new discoveries and more fundamental information?
3. Be
critical - could the authors have used better controls, more modern techniques,
more definitive experimental approaches?
Bibliography:
1. Be
consistent; include all authors, title of article, source, year; refer
to articles in text either by number or by authors and year.
Overall
presentation of paper that will be evaluated:
1. Have
you followed the above outline and included what is requested above?
2. Depth
of coverage?
3. Clarity?
4. Sentence
structure, spelling, grammar, and physical appearance.
Evaluation
of Biol551 Paper
(Attach
to Paper)
Student
Name________________ Grade
(out of 100 possible points) ______
Introduction:
Definition
of problems and issues to be discussed: (10 POINTS)
Presentation
of background information and previous results: (10 POINTS)
Evaluation
of research in paper:
Questions
being addressed: (10 POINTS)
Description
and evaluation of techniques used: (10 POINTS)
Description
of current research discoveries: (10 POINTS)
Discussion:
Adequate
evaluation of work presented: (10 POINTS)
Description
of questions that remain to be answered, directions most likely to lead
to new discoveries: (10 POINTS)
Bibliography: Inclusion
of all authors, title of article, source, year; consistency: (5 POINTS)
Overall
presentation of paper:
Depth
of coverage: (10 POINTS)
Clarity:
(5 POINTS)
Sentence
structure, spelling, grammar, and physical appearance: (10 POINTS)