Production of Protein Kinases in E. coli
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 wordsBacterial protein expression Protein purification Kinase Aurora-A
This work is supported by an Imperial College Junior Research Fellowship and grant BB/M021149/1 from the BBSRC.
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