Production of Protein Kinases in E. coli

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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 languageEnglish
Title of host publicationHeterologous Gene Expression in E. coli
EditorsN. Burgess-Brown
Place of PublicationNew York
PublisherSpringer
Pages251-264
Number of pages14
Volume1586
ISBN (Print)978-1-4939-6885-5
DOIs
Publication statusPublished - 2017

Publication series

NameMethods in Molecular Biology
PublisherHumana Press, New York
Volume1586

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  • Cite this

    Dodson, C. (2017). Production of Protein Kinases in E. coli. In N. Burgess-Brown (Ed.), Heterologous Gene Expression in E. coli (Vol. 1586, pp. 251-264). (Methods in Molecular Biology; Vol. 1586). Springer. https://doi.org/10.1007/978-1-4939-6887-9_16