[PEP] Protein Expression and Purification

[PEP] Protein Expression and Purification

Protein expression and purification is an essential part structural biology. This module will introduce the theory and practice of protein expression and purification. Students will have the opportunity to use modern, robotic protein purification equipment.

Lecturer

Dr Arvind Varsani (Electron Microscope Unit) has a Pharmaceutical Chemistry and Molecular Biology background. He will present lectures on various aspects of virus structures, assist in the electron microscopy module and provide molecular biology support for the programme.

Main Outcomes

The ability to:

  • describe and discuss protein expression and purification.
  • express recombinant proteins
  • purify proteins by ion exchange chromatography, gel filtration chromatography, affinity chromatography and ultracentrifugation.

This module descriptor document also can be downloaded as an MS-Word document.

Main Content

  • Expression systems
  • Introduction to protein purification
  • Detection of proteins
  • Principles of chromatography (ion exchange, gel filtration, affinity chromatography
  • Centrifugation
Home Department: Molecular and Cell Biology, UCT
Module description (Header): Protein expression and purification
Generic module name: Structural Biology
Alpha-numeric code: STB705
Credit Value: 5 Credits
Duration: 3 Weeks
Module Type: P
Level: 8
Prerequisites: None
Co-requisites: None
Prohibited combinations: None
Learning time breakdown (hours):
Contact with lecturer/tutor: 55
Assignments & tasks: 15
Tests & examinations: 0
Practicals: 20
Selfstudy: 10
Total Learning Time 100
Methods of Student Assessment: Students will be required to submit a practical write up and complete an assignment.

Assignments – 100%

Assessment

Students will be required to submit a practical write up and complete an assignment (100% of course mark).

Online Lectures

  • [PEP] Overview Lecture by Arvind Varsani
  • Recombinant Expression Systems
  • Molecular Biology and Gene Cloning for the Structural Biologist - by Michael Benedik, Department of Biology, Texas A&M University
    See /lectures/benedik/ on the server.

    • Introduction: Genes and Gene Expression
    • Cloning DNA
    • DNA Synthesis, PCR, DNA Sequencing
    • Mutagenesis and Protein Engineering: mutilation and mayhem as applied to poor defenseless genes.
    • Gene expression in prokaryotes
    • Gene expression in yeast and eukaryotes
  • The BirkBeck College protein purification course

Documentation and Resources

See the following resources on our website, for further protocols and information, including:

References

Amersham Pharmacia Biotech. 2000. Antibody Purification: Handbook. Sweden: Amersham Pharmacia Biotech.

Baneyx, F. 1999. "Recombinant protein expression in Escherichia coli." Current Opinion in Biotechnology, 10 (5): 411-421.

Cereghino, GPL and JM Cregg. 1999. "Applications of yeast in biotechnology: protein production and genetic analysis." Current Opinion in Biotechnology, 10 (5): 422-427.

Geisbrecht, BV, S Bouyain, and M Pop. 2006. "An optimized system for expression and purification of secreted bacterial proteins." Protein Expression and Purification, 46 (1): 23-32.

Guruatma, JK. "Agarose Gel Electrophoresis. ASU - Ask A Biologist." 2010. http://askabiologist.asu.edu/agarose-gel-electroporesis (accessed 2013-02-05).

Guruatma, JK. "Alkaline Lysis. ASU - Ask A Biologist." 2010. http://askabiologist.asu.edu/alkaline-lysis (accessed 2014-04-16).