Construction and identification of human Angiopoietin2 prokaryotic expression vector

Abstract

Objective To construct the human Angiopoietin2 prokaryotic expression vector.Methods Complete Angiopoietin2 gene was cloned by reverse transcription-polymerase chain reaction (RTPCR) fromin vitro cultured human umbilical vein endothelial cells (HUVECs),then the gene was cloned into pET32a plasmid expression system,and ultimately Angiopoietin2 was stablely expressed in E.coli BL21.Angiopoietin2 protein was then purified and identified by condensate gel electrophoresis,SDS-PAGE and gene sequencing.Results The cloned Angiopoietin2 gene fragment from HUVECs was about 1.5 Kb,the Angiopoietin2 protein was expressed by pET32a plasmid expression system,and it was predicated about 70 kDa protein by SDS-PAGE.Conclusion The prokaryotic expression vector of Angiopoietin2 is successfully constructed by recombinant DNA techniques. Key words: Angiopoietin; Prokaryotic expression vector