Survival protein recombinant adenovirus vector and its expression in dendritic cells.docVIP

 PAGE \* MERGEFORMAT 24 Survival protein recombinant adenovirus vector and its expression in dendritic cells Author: Xiong-peng, CHEN Zhi-zhe, Lin Xu, Hu Da, Li Chun group, Ting Yang, XU Zhen-books, Lvlu Lu, Chen Cai-Ping, Zhang Lang-hui Abstract This study aimed to construct containing human survivin (survivin) gene recombinant adenovirus vector, and to explore its in transfected dendritic cells. Plasmid pcDNA3.0-survivin as a template obtained by PCR amplification of full-length sequence of survivin gene. PCR products recovered by digestion, and directed into adenovirus shuttle plasmid to obtain recombinant plasmid pShuttle-CMV-survivin. By double digestion, PCR and sequencing inserts, will be the correct recombinant pShuttle-CMV-survivin into E.coli BJ5183 bacteria (including the adenovirus backbone plasmid) and homologous recombination, and screening positive clones, extract quality tablets, for this recombinant plasmids were digested, Linearization, purification, LipofectamineTM 2000 with liposome-mediated transfection of 293 cells. Preparation of viral supernatant and determine its titer, the virus supernatant transfected dendritic cells, the application method of Western blot analysis of survivin expression. The results showed that: The successful construction of recombinant adenovirus containing human survivin gene, the virus titer: 2.65 × 109pfu/ml. Identification of Western blot showed that DC transfected by the recombinant adenovirus can be effectively expressed survivin. Conclusion: The recombinant adenovirus vector containing the survivin gene was successfully constructed for further studies to carry out anti-leukemic immune to lay an experimental basis. Keywords: adenovirus vector; survival protein gene; dendritic cells Construction of Recombinant Adenovirus Vector Containing Human Survivin Gene and Its Expression in Dentritic Cells Abstract The study was aimed to construct the recombinant adenovirus vectors contai