Gene cloning, expression and homology modeling of e6 gene from human papillomavirus 16

These high risk HPV types, usually HPV-16 can be found in over 80% of cervical carcinomas. E6 is a significant toxin secreted by HPV-16, which contributes pathogenicity of to women. The complete ORF of E6 gene (2482 bp) was amplified using PCR. It was cloned in TA and sub-cloned in pET28a vector then transformed into Escherichia coli BL21(DE3) codon plus RP cells expressed by the induction with 1.0 mM of IPTG. The expected size of expressed protein was 68.0 kDa estimated by migration in 12% SDS–PAGE. Anti-His monoclonal antibodies were used to sub- stantiate the recombinant protein by Western blotting. The percent similarity between E6 of HPV-16 with other HPVs E6 toxins revealed that the E6 sequence varied from 99.35 to 50.40%. Homology modeling was used to construct 3-D structure of E6 of HPV-16 with the known crystal 3-D structure (PDB: 1PRE). This protein can be used for immunoassays and it is suitable for vaccine candidate against HPV-16 infection.