Prokaryotic Expression of HPV18 E2

Objective Construction of Prokaryotic Expression vector,pET28a(+)-HPV18E2 expressing human papillomavirus(HPV) 18 E2 gene.Prokaryotic expression and purification of HPV18 E2 protein,for future study of HPV18 E2 vaccine against HPV related cervical cancer.Methods 1) Amplification of HPV18 E2 gene cDNA by PCR from HPV 18 genomic DNA template,insert of E2 cDNA fragment into prokaryotic expression vector pET28a(+),to construct recombinant prokaryotic expression vector pET28a(+)-HPV18E2;2) Confirmation of the recombinant prokaryotic expression vector by PCR,restrictional enzymes digestion and DNA sequencing;3) Transformation of BL21 E.coli with pET28a(+)-HPV18E2.Expression of HPV18 E2 protein by IPTG induction and protein purification with Ni-NTA gel chromatography column;4) E2 protein expression was verified by SDS-PAGE and Western blotting.Results 1) The construction of recombinant plasmid was confirmed by PCR,double enzymes digestion and DNA sequencing.2) Expression of HPV E2 protein was purified and analyzed by SDS-PAGE and Western blotting assay and it was shown that the protein prokaryotic expressed was corrected and purified.Conclusions HPV18 E2 protein was prokaryotic expressed and purified,and would be used for future study of vaccine against HPV related cervical cancer.