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Production of Protein Kinases in E. coli

  • Charlotte A. Dodson
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1586)

Abstract

Recombinant protein expression is widely used to generate milligram quantities of protein kinases for crystallographic, enzymatic, or other biophysical assays in vitro. Expression in E. coli is fast, cheap, and reliable. Here I present a detailed protocol for the production of human Aurora-A kinase. I begin with transformation of a suitable plasmid into an expression strain of E. coli, followed by growth and harvesting of bacterial cell cultures. Finally, I describe the purification of Aurora-A to homogeneity using immobilized metal affinity and size exclusion chromatographies.

Key words

Bacterial protein expression Protein purification Kinase Aurora-A 

Notes

Acknowledgments

This work is supported by an Imperial College Junior Research Fellowship and grant BB/M021149/1 from the BBSRC.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Molecular Medicine, National Heart & Lung InstituteImperial College LondonLondonUK

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