Cloning and Prokaryotic Expression of Human CD271 Gene

Objective To clone human CD271 gene and express in E. coli. Methods Amplify CD271 gene from human REH cell line by RT-PCR and clone into expression vector pET22b. Transform the constructed recombinant plasmid pET22b / CD271 to E. coli BL21(DE3) for expression under induction of IPTG. Purify the expressed product by affinity chromatography and identify by Western blot. Results The restriction analysis of recombinant plasmid pET22b / CD271 showed a target gene fragment at length of 1197 bp. SDS-PAGE showed that the relative molecular mass of expressed protein was about 75 000. The expression level of target protein reached a peak value under induction of 0. 5 mmol / L IPTG. The purified target protein reached a purity of 82. 5% and showed good reactogenicity as proved by Western blot. Conclusion Human CD271 gene was successfully cloned and expressed in E. coli.