Expression,purification and preliminary identification TGF β1 in E.coli

Objective To was express TGF β1 in prokaryotic vector pET 43.1 for production.Methods TGF β1 gene synthesized with enterokinase(EK)digested sequence,the gene was cloned into the NusA protein fusion expression vector pET 43.1.Then the recombinant vector was introduced into E.coli BL21(+) for expression which was induced by IPTG.Fusion protein was digested by EK and purified by Ni-affinity chromatography.Results SDS-PAGE result showed that the expression level of recombinant protein was up to about 80%,and most of them were soluble.The purity of target protein was about 95% after Ni-affinity chromatography.TGF β1 was harvested with a recovery of 80% by EK digestion then.The result showed that TGF β1 has strong immunogenicity against TGF β1 antibody.Conclusions Recombinant TGF β1 is produced with high yield in NusA-pET 43.1 system by E.coli BL21(+).