The Biotechnology Curriculum Collection of the California Community Colleges

ORANGE COAST COLLEGE COURSE OUTLINE OF RECORD

Course developer: Farah Sogo
Course static ID:
TOP No. 0430.00
CIP No.
Course adoption:
Date revised:
Revised by:
Semester(s) offered: Spring
Distance education alternative

COURSE MASTER DICTIONARY DATA

Title 5 credit status: Associate degree credit course X
Nondegree credit course
Noncredit course
Course name/number: Biology 251
Division: Math and Science
Course title: Advanced Laboratory Methods in Biotechnology
Department: Biology
Units: 5
Total course hours: 162
Course length: 16 weeks
Weekly hours configuration: 3.0 hours lecture 6.0 hours lab
Grading method: Graded X
CR/NC
Student option
Noncredit
Method of Instruction: 30 (2 digit no.)
Basic skills status: N (P, B or N)
Materials fee: No
Yes X
$ 12

Justification: X-ray and thermal film, nylon membranes, and gels are used to analyze DNA and protein, and for permanent reference and technique records.

COURSE PREREQUISITE:
Biology 250

CATALOG DESCRIPTION:
This course will simulate a research laboratory setting and industrial setting with weekly lab meetings. Laboratory experiences will include plant and animal tissue culture, RNA and DNA isolation, genomic library construction and analysis, and PCR. Lecture will primarily focus on theory behind the uses of biotechnology in a research laboratory and industrial settings.

SCHEDULE DESCRIPTION:
An advanced laboratory-based course which introduces modern biotechnological techniques including tissue culture, RNA and DNA isolation, genomic library constructions, and PCR.

COURSE CLASSIFICATION:

A Liberal arts/AA X
B Remedial
C Remedial
D-H Community course
I Occupational required
I Occupational elective

COURSE TRANSFER:

0 Non-transfer/Non-AA
1 Non-transfer AA
2 Transfer CSU X
3 Transfer UC


JUSTIFICATION FOR THE COURSE:
Required for certification
Occupational state-of-the-art
Advisory committee recommendation

COURSE CONTENT AND SCOPE/TOPIC OUTLINE:

LECTURE OUTLINE

1) Manipulation of Gene Expression in Prokaryotes

a) Regulatable promoters
b) Large scale protein production

i) Fusion proteins
ii) Immunoaffinity chromatographic purification
iii) Translation Expression vectors

2) Translation review
3) Recombinant Protein Production in Eukaryotic cells

a) Mammalian Cell Expression vectors
b) Insect Cell Expression systems
c) Plant Genetic Engineering

4) Directed Mutagenesis

a) Oligonucleotide-Directed Mutagenesis

i) With M13 DNA
ii) With plasmid DNA
iii) PCE-amplification
iv) With degenerate oligonucleotides

5) Protein Engineering

a) Adding disulfide bonds
b) Altering amino acids
c) Reducing free sulfhydryls
d) Modifying Enzyme specificity

6) Monoclonal Antibodies

a) Structure an function of antibodies
b) Chemically linked monoclonal antibodies
c) Hybrid Cell production

i) Hybrid cell line selection and identification

d) Producing monoclonal antibodies in E. coli

7) Microbial Production of Therapeutic Agents

a) Pharmaceuticals

i) Isolation of interferon cDNAs and engineering human interferons
ii) Engineering Human Growth Hormone
iii) HIV Therapeutic Agents

8) Vaccines

a) Subunit Vaccines (purified and recombinant protein antigens)

i) Herpes Simplex Virus
ii) Peptide Vaccines
iii) Genetic Immunization

b) Vector Vaccines

i) Anti virus
ii) Anti bacteria

9) Synthesis of Commercial Products by Recombinant Microorganism

a) Antibiotics

i) Cloning Antibiotic Biosynthesis genes
ii) Synthesis of Novel antibiotics
iii) Improving production of antibiotics

b) Biopolymers

i) Xanthan Gum production
ii) Melanin biosynthesis
iii) Rubber

10) Large-Scale Production of Proteins from Recombinant Microorganisms

a) Microbial growth principals

i) Batch fermentation
ii) Continuous fermentation
iii) Maximizing efficiency

b) Bioreactors
c) Processing proteins

i) Harvesting cells
ii) Disrupting Cells
iii) Protein solubilization

11) Regulating the Use of Biotechnology

a) Regulating food and food ingredients

i) Code of regulations

b) Release of genetically engineered organisms
c) Human Gene Therapy

LABORATORY OUTLINE

1) Animal Tissue Culture and Sterile Techniques
2) RNA and DNA isolation from animal cultured cells
3) Plant Tissue Culture and Sterile Technique
4) RNA and DNA isolation from cultured plant cells
5) Restriction Mapping of the Lambda Chromosome
6) Genomic Library Construction and analysis

a) Construction of a genomic library of l DNA
b) Transformation of E. coli with l Library
c) Colony Hybridization of the l Library
d) Purification and Identification of l Clones

7) Cloning by Polymerase Chain reaction

a) Amplification and Purification of a l DNA fragment
b) Recombination of a PCR Product and Plasmid pBLU
c) Transformation of E. coli with PCR Product
d) The Purification and Identification of PCR/pBLU Recombinants

8) Oligonucleotide directed mutagenesis to insert a novel restriction site into the LEU2 gene.
9) IPTG induction of recombinant b-galactosidase in E. coli
10) Collaborative Project with Biology 221 students: The biology 251 students create primary tissue culture from heart chick cells. The cells will be given to the biology 221 students for microscopic analysis.
11) Collaborative Project with Biology 100 students: The Biology 100 students each prepare a sample of their DNA for PCR analysis. The Biology 251 will use PCR to detect Alu Insertion Polymorphisms and VNTR Polymorphisms. The DNA results will be photographically documented and returned to the Biology 100 students. The students will be involved in preparing a video presentation that explains the lab procedures and results to the Biology 100 class to ensure uniformity of concept presentation to all Biology 100 laboratory sections.
12) Collaborative Project with Biology 125: The biology 251 students will prepare a poster session demonstrating equipment and techniques they have used throughout the semester as well as examples of data generated using the equipment. The Biology 125 student classes will have an opportunity to come to the poster session during there regularly scheduled class period. Since Biology 125 has many sections, Biology 251 students will sign up for a time slot to present the posters.
13) Collaborative Project with Biology 225: The Biology 225 students will prepare protein samples from their own blood. The proteins will be analyzed for hemoglobin by electrophoresis by the Biology 251 students. The results will be documented and returned to the Biology 225 students along with an oral presentation.

INSTRUCTIONAL OBJECTIVES:

The student will be able to:

1. Maintain sterile tissue culture
2. Isolate RNA and DNA from target tissues with minimal degradation.
3. Construct a genomic library from phage l DNA
4. Clone a l DNA fragment using PCR and purify the DNA product.
5. Prepare meticulous data sheets derived from sample analysis for inspection by collaborating biology classes.
6. Prepare and present a scientific poster.
7. Describe how protein can be produced in prokaryotes and eukaryotic cells.
8. Describe Oligonucleotide-directed mutagenesis and the industrial applications of this technique.
9. Describe how to engineer protein.
10. Describe how monoclonal antibodies are produced and used.
11. Describe how recombinant microorganisms are used to produce pharmaceuticals, vaccines, biopolymers and antibiotics in a large-scale industrial setting.
12. Be familiar with the current regulation regarding the use of biotechnology in food, humans and genetically engineered organisms.
13. Maintain a laboratory notebook.
14. Conduct themselves in mock "meetings" in a professional fashion.

METHOD OF STUDENT EVALUATION:
Student written exams, practical exams, maintenance of a laboratory notebook, present a scientific poster, production of data sheets for Biology 100 students and Biology 225 students.

INSTRUCTIONAL METHODOLOGIES:
Lecture on key concepts
Collaborative projects with other biology classes (Biol 100, 221, 125, 225) in which the students send samples to the Biology 250 students for further analysis and data sheet production, create products, and offer a poster session.
Lab meetings that simulate a research lab meeting and/or a production meeting in industry.
Computer analysis of data using databases available on the internet.

WRITING ASSIGNMENTS/PROFICIENCY DEMONSTRATION:
Students will successfully complete entries on a daily basis into a laboratory book.
Completion of data sheets and written summaries of data provided by collaborative sample analyses.

REPEATABILITY:

N/A

FEASIBILITY:

Faculty: This course will be taught using existing faculty.
Classroom: This course will be taught in existing classroom facilities
Library Learning Resources: The library has an interlibrary loan program and internet access that will be sufficient any necessary classroom research.

EDUCATIONAL MATERIALS:

Molecular Biotechnology: Principles and Applications of Recombinant DNA, Glick, B.R. and Pasternak, J.J., ASM Press, Washington, D.C.
Laboratory DNA Science, Bloom, M., Freyer, G., Micklos, D., Benjamin/Cummings Publishing Company, Inc.