Gene Cloning and Eukaryotic Expression of HPV 1611 Gene
Abstract: To clone and express in vitro Human papillomavirus type 16 l1 (HPV16L1) gene ,and provide a good basis for further research DNA vaccine against HPV16 infection and human cervical cancer, the HPV16l1 gene fragment was amplified from genome of p16L1BN1 by PCR and cloned into eukaryotic expression vector pcDNA3.0.Sequencing showed the plasmid pcDNA3-HPV16L1 was constructed successfully. Then the recombinant plasmid pcDNA3-HPV16L1 transfected 7721 human liver cancer cells using Lipofectamine 2000 reagent. The molecular weight of the expressed protein, was 55kDa, analyzing by SDS-PAGE, as same as the molecular weight of HPV16L1. The Western blotting result show that the expressed protein could be detected by HPV16L1 specific monoclonal antibody. This study provided a good basis for further research on HPV16L1 DNA vaccine.