Construction of recombinant plasmid pQE32-HPV18L1 and protein expression

Objective To construct a recombinant prokaryotic expression vector for obtaining the HPV18L1 protein and develop the genetic engineering vaccine.Methods The L1 gene of HPV18 was amplified by PCR from DBR322-HPV18 and cloned into pUC19.The sequence of cloned HPV18L1 was confirmed by restriction analysis and DNA sequencing.A HPV18L1 prokaryotic expression plasmid,pQE32-HPV18L1,was then constructed by subclone.pQE32-HPV18L1 tranformed E.coli M15 was induced by IPTG.The expression of L1 protein was analyzed by SDS-PAGE and western blot.Results The amplified DNA fragment was in size of 1.7Kb as expected.Restriction analysis showed that the amplified gene was inserted in pUC19 correctly.Sequence showed there was no mutation in both ends of the cloned L1.Restriction analysis showed that the recombinant pQE32-HPV18L1 right.A 63KD protein could be seen in M15 induced by IPTG with SDS-PAGE and was positive while reacting with HPV18L1 antibody by western blot.Conclusion HPV18L1 was successfully cloned and expressed in this study.