Production of Protein Kinases in E. coli

Charlotte Dodson

doi:10.1007/978-1-4939-6887-9_16

Production of Protein Kinases in E. coli

Charlotte Dodson

Research output: Chapter or section in a book/report/conference proceeding › Chapter or section

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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.

Original language	English
Title of host publication	Heterologous Gene Expression in E. coli
Editors	N. Burgess-Brown
Place of Publication	New York
Publisher	Springer
Pages	251-264
Number of pages	14
Volume	1586
ISBN (Print)	978-1-4939-6885-5
DOIs	https://doi.org/10.1007/978-1-4939-6887-9_16
Publication status	Published - 31 Dec 2017

Publication series

Name	Methods in Molecular Biology
Publisher	Humana Press, New York
Volume	1586

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10.1007/978-1-4939-6887-9_16

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Production of Protein Kinases in E. coli

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