Construction And Identification Of Prokaryotic Expression System With L1 Gene Of Human Papillomavirus Type 11

Recently, sexually transmitted diseases (STDs) have been annuallyincreased. Condyloma Acuminatum (CA), the second cause of STDs isthe pathogen of 24.73% cases of STDs. Human papillomavirus (HPV) iscomposed of various serotypes and CA is mainly caused by HPV11. Thelate gene L1 of HPV genome encodes the major capsid protein, whichcontains stable immunogenicity. In the prokaryotic expression system,L1 gene may express and automatically assemble virus-like particles(VLPs) lack of viral DNA. VLPs possess the dimensional conformation,immunological characteristics and biological properties similar to wildHPVs. Subsequently, vaccines composed of the protective antigen L1may play an important role in the prevention of HPV infections.Our study firstly got HPV DNA from fresh tissue samples of CA fromTianjin area and then typified them with PCR to screen the HPV11 DNA.The L1 gene of HPV11 with enzymatic identification sites was multipliedwith PCR. Plasmid pMD18T-HPV11L1 was constructed by basiccomplementary linkage between the single basic A at the both end of thePCR products and the single basic T at the end of vector pMD 18-T,which was then identified by enzymic digestion, PCR of L1 gene as wellas its sequence analysis. L1 gene digested from pMD18T-HPV11L1 wasinserted into the expression vector plasmid pET42a with the same cohesive ends to construct the recombinant expression plasmidpET42a-HPV11L1 expressing HPV11L1 protein, which was thensuccessfully identified by enzymic digestion PCR of L1 gene and itssequence analysis. Finally, the recombinant expression plasmidpET42a-HPV11L1 was primarily expressed in the E. coli strain BL21(DE3).Our study successfully constructed pET42a-HPV11L1 by cloning L1gene of HPV11 into the prokaryotic expression vector pET42a and thengot high expression of L1 gene in E. coli, which created conditions forfurther study of HPV vaccine.