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Biomolecular and Bioanalytical Techniques
book

Biomolecular and Bioanalytical Techniques

by Vasudevan Ramesh
June 2019
Beginner to intermediate content levelBeginner to intermediate
576 pages
25h 29m
English
Wiley
Content preview from Biomolecular and Bioanalytical Techniques

6 Overproduction, Separation and Purification of Affinity‐Tagged Proteins from Escherichia coli

Finbarr Hayes1 and Daniela Barillà2

1 Faculty of Biology, Medicine and Health, The University of Manchester, Manchester, M13 9PL, UK

2 Department of Biology, University of York, York, YO10 5DD, UK

6.1 Introduction

Understanding protein function is a central goal of modern molecular biology. This aim typically is achieved using a combination of in vivo and in vitro approaches that together provide an integrated overview of protein features, interactions and regulation. The availability of a sufficient quantity of pure protein that can be isolated rapidly and efficiently is crucial for its biochemical, physical and structural characterisation in vitro. Moreover, in vitro studies of purified proteins are key to the development of drugs that affect proteins implicated in disease formation and progression, as well as to the optimization of proteins produced for biotechnological, bioprocessing and other industrial and commercial purposes.

The Gram‐negative bacterium Escherichia coli remains the primary cell factory for protein production: approximately 90% of the tertiary structures that have been deposited in the Protein Data Bank have been generated from proteins that were purified from E. coli [1]. E. coli is specially suitable for protein production due to the bacterium's rapid growth kinetics in inexpensive cultivation media, high cell density, simplicity of handling ...

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ISBN: 9781119483960Purchase book