Construction and identification of recombinant plasmid pQE32-HPV18 L1

Objective To construct a recombinant plasmid pQE32 HPV18 L1 for obtaining the HPV18 L1 protein expressed in E.coli and developing the genetically engineered vaccine.Methods PCR was used to amplify the HPV18 L1 gene from plasmid pBR322 HPV18 in which the HPV18 L1 was cloned. A new plasmid, pUC19 HPV18 L1, was then constructed by inserting the amplified HPV18 L1 gene into pUC19, a prokaryotic expression vector.Restriction analysis and sequencing were used to confirm the structure of pUC19 HPV18 L1.A new plasmid, pQE32 HPV18 L1, was then constructed by inserting the amplified HPV18 L1 gene into pQE32 a prokaryotic expression vector.Results A DNA fragment in size of 1.7Kb was amplified. Restriction analysis showed that the amplified gene was inserted into pUC19 correctly. Sequencing showed there was no mutation in both ends of the inserted L1 gene.Conclusion The plasmid pQE32 HPV18 L1 is constructed successfully.