Construction of HPV16E6 Gene Plasmid Expressing in Prokaryotic System

The objective of this paper is to build HPV16 E6 gene expression plasmid pET28a(+)-HPV 16 E6.The genomic DNA extracted from CaSki cells are employed as a template,a specific pair of primers is designed,HPV16E6 gene fragment is amplified by PCR.HPV16E6 gene fragment and plasmid pET28a(+) digested by NcoⅠand Hind Ⅲ are connected by T4 DNA ligase.Results show that the construction of recombinant plasmid pET28a(+)-HPV16 E6 is transferred into E.coli DH5α,the positive clones are screened by PCR and enzyme digestion,which are transferred into E.coli BL21 DE3 Star Plyss,HPV16E6 gene expression plasmid pET28a(+)HPV16E6 are built successfully.It is concluded that HPV16 E6 gene expression plasmid pET28a(+)-HPV 16 E6 is ready to prepare HPV16E6,which provides a foundation of the early diagnosis for HPV16 in cervical tissue and therapeutic vaccine about HPV16.