Prokaryotic expression and purification of CDK1

The CDK1 cDNA sequence (GenBank: {"type":"entrez-nucleotide","attrs":{"text":"XM_005270303.3","term_id":"1034570512","term_text":"XM_005270303.3"}}XM_005270303.3) was synthesized by Sangon Biotech. Recombinant cDNA was further cloned into BamH I and Hind III sites of pET-28a(+) (Invitrogen, Frederick, MD, USA) to generate the prokaryotic expression vector pET28-CDK1. Thereafter, vector was transformed into Escherichia coli BL21 (TransGen Biotech, Beijing, China) to express recombinant protein CDK1 at 25°C. The recombinant protein CDK1 was purified by Ni-column affinity chromatography (Jingcheng, Wuhan, China).