HPV18E2 protein CTL epitope peptides Construction and Expression of.docVIP

 PAGE \* MERGEFORMAT 12 HPV18E2 protein CTL epitope peptides Construction and Expression of [Abstract] Objective To analyze the HPV18E2 protein CTL epitopes, synthesized and purified the epitope peptide, vector expressing HPV18E2 protein target cells to test-specific CTL on target cell-killing effect, for the further development of human papillomavirus type 18 (HPV18 ) lay the foundation for genetic engineering vaccine. Methods The recombinant plasmid (pBR322  HPV18) as a template, using HPV18E2 DNA fragment was amplified by PCR, the HPV18E2 DNA with the strengthening of the green fluorescent plasmid (pIRES2  EGFP) recombinant plasmid (pIRES2  HPV18E2  EGFP). Check with the enzyme electrophoresis and sequencing of plasmid sequences after the reorganization correctness. Hela cells transfected with recombinant plasmid. Experimental assessment of 51Cr release CTL killing capacity. Results Cloning of recombinant plasmid pIRES2  HPV18E2  EGFP displayed after restriction endonuclease digestion with the same expectations, and the sequence verified insert sequence does not change. Transfection and selection with G418 after the fluorescence microscope can be seen the expression of green fluorescent cells. Three candidate peptide-specific anti-personnel capacities of 60.00%, 71.29% and 80.00%. Conclusion three alternative peptides have significant anti-effect, are effective HPV18E2 protein CTL epitopes. [Keywords:] human papillomavirus type 18; E2 gene; peptide Construction and expression of recombinant plasmid pIRES2  HPV18  EGFP Abstract: Objective To analyse the function of epitope  specifical HPV18E2 CTL. Methods The E2 gene of HPV18 was amplified by PCR from pBR322  HPV18 and cloned into pIRES2  EGFP. The sequence of cloned HPV18E2 was confirmed by restriction analysis and DNA sequencing. Then transfected into HeLa cells. The functional analysis is valued by 51Cr  release assay. Results HPV18 E2 gene fragment was amplified by PCR and li