Human papillomavirus 16 L1-CEA CTL epitope fusion protein in prokaryotic expression.docVIP

 PAGE \* MERGEFORMAT 13 Human papillomavirus 16 L1/CEA CTL epitope fusion protein in prokaryotic expression Author: Zhang Italian ZHU Guan-Bao, ZHANG Li-fang, Zhu Shan Li, Chen Jun [Abstract] Objective: To construct human papillomavirus (HPV) 16 type L1 capsid protein and carcinoembryonic antigen (CEA) cytotoxic T cells (CTL) epitope chimeric gene, to explore its expression in prokaryotic expression systems. Methods: CEA CTL epitopes (CEA691 IMIGVLVGV), with HPV16-positive cervical cancer extracted DNA as a template by PCR amplified HPV16 L1/CEA CTL epitope chimeric gene was cloned into pET32a () expression vector, transformation E. coli BL21 (DE3), fusion protein was expressed by IPTG induction, through the Ni-NAT ion-exchange chromatography and purified further use of SDSand Western-blot analysis of identification. Results: pET32a () / HPV16 L1/CEA CTL epitopes in the recombinant plasmid was successfully constructed, and were expressed in E. coli. SDSanalysis showed that the expression product molecular weight (Mr) of approximately 83 * 103, in line with expectations, Western-blot method to further confirm it as the target protein. Conclusion: The use of pET32a () expression vector to express HPV16 L1/CEA CTL epitope recombinant protein for further study of immune effects of laying a foundation. [Keywords:] papilloma virus; carcinoembryonic antigen; prokaryotic expression; T-lymphocytes, cytotoxic Abstract: Objective: To construct the chimeric gene of HPV16 capsid protein L1 and CEA CTL epitope and to explore the protein expression of it in Escherichia coli. Methods: The CEA CTL epitope (CEA691 IMIGVLVGV) was selected and the chimeric gene of HPV16 L1 / CEA CTL epitope was amplified by PCR. The chimeric gene was cloned into the expression vector pET32a () for forming the pET32a () / HPV16 L1/CEA CTL epitope recombinant with which is transformed E.coli BL21. The pET32a () / HPV16 L1 / CEA CTL epitope fusion protein was expressed in E.coli, induced by IPT