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 in Book/Report/Conference proceeding › Other chapter contribution

2 Citations (Scopus)

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 - 2017

Publication series

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

Access to Document

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

Fingerprint Dive into the research topics of 'Production of Protein Kinases in E. coli'. Together they form a unique fingerprint.

Cite this

@inbook{27f58ca352654a5ba554662147f364a4,

title = "Production of Protein Kinases in E. coli",

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.",

author = "Charlotte Dodson",

year = "2017",

doi = "10.1007/978-1-4939-6887-9_16",

language = "English",

isbn = "978-1-4939-6885-5",

volume = "1586",

series = "Methods in Molecular Biology",

publisher = "Springer",

pages = "251--264",

editor = "N. Burgess-Brown",

booktitle = "Heterologous Gene Expression in E. coli",

}

TY - CHAP

T1 - Production of Protein Kinases in E. coli

AU - Dodson, Charlotte

PY - 2017

Y1 - 2017

N2 - 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.

AB - 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.

U2 - 10.1007/978-1-4939-6887-9_16

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

M3 - Other chapter contribution

SN - 978-1-4939-6885-5

VL - 1586

T3 - Methods in Molecular Biology

SP - 251

EP - 264

BT - Heterologous Gene Expression in E. coli

A2 - Burgess-Brown, N.

PB - Springer

CY - New York

ER -