Abstract:

Abstract:Objective To construct prokaryotic expression vector harboring human papillomavirus-16 E7(HPV-16) gene and   observe the E7 protein expression.Methods Synthetic E7 gene was amplified using polymerase chain reaction and subcloned into prokaryotic expressing vector pGEX-4T1.The recombinant plasmids,pGEX-4T1-HP
V16-E7,were transferred into engineering bacteria Rosetta,and the fusion proteins E7-GST were analyzed on SDS-PAGE and purified by　Ni2+-NTA affinity chromatography under induction with IPTG.Results The target gene fragment at a length of 300 bp was observed on the agarose gel electrophoresis pattern and PCR.SDS-PAGE analysis showed that the expressed recombinant protein,with a relative molecular mass of about 36 000,could be found.Conclusion The recombinant protein,E7-GST,is effectively expressed in prokaryotic expression system.

Key words: human papillomavirus-16；polymerase chain reaction；prokaryotic expression vector