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 | |
| Publication status | Published - 2017 |
Publication series
| Name | Methods in Molecular Biology |
|---|---|
| Publisher | Humana Press, New York |
| Volume | 1586 |
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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