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Protein Expression, Purification and Biophysical Characterisation

Overview

  • Credit value: 30 credits at Level 7
  • Convenor and tutor: Professor Nicholas Keep
  • Assessment: a 1.5-hour examination (66%) and coursework (34%)

Module description

Producing milligram amounts of pure protein is a necessary precursor to all structural biology techniques. This module explains the theory of how to do this, how to characterise the protein sample and offers possible solutions to problems encountered in achieving good protein expression.

Indicative module syllabus

  • Manipulation of DNA
  • High Level of Expression of Proteins in E.coli
  • Trouble Shooting Protein Expression and Expression of Complexes
  • Protein Purification
  • Protein Characterisation
  • Other Expression Hosts

Learning objectives

By the end of this module, you will be able to:

  • enumerate and explain the steps involved in expressing a protein for structural studies in a bacterial host
  • list other possible hosts for protein expression and briefly explain their application
  • troubleshoot problems in protein expression
  • give examples of common methods of protein purification and explain how each exploits particular physicochemical properties of proteins in achieving separations
  • list and explain common techniques employed to characterise a purified protein sample.