Prokaryotic Expression and Purification of Recombinant Zebrafish CD36 Protein

Objective: To construct prokaryotic expression vector of recombinant zebrafish CD36 protein,and to purify the protein of extracellular amino acid residues 38 to 432 segments.Methods: The coding sequence of ze brafish CD36 protein was amplified by PCR and inserted into the prokaryotic expression vector pET-28a with 6× His tag to construct the recombinant plasmid pET28a-CD36.The recombinant plasmid was transformed into E.coli BL21(DE3),and the expression of fusion protein was induced by IPTG.Ni2 + metal chelating column was utilized for the purification of the fusion protein after the expression conditions were optimized.Results: Recombinant plas mid pET28a-CD36 was constructed and the recombinant protein was expressed in E.coli successfully.After being purified by affinity chromatography,SDS-PAGE showed a clear protein band with a relative molecular weight of 46.8 kD expectedly.Conclusion: The fusion protein of zebrafish CD36 was successfully expressed and purified,which lays the foundation of further study on its biological function.