Human bax gene retroviral expression vector and its expression in Hela cells.docVIP

 PAGE \* MERGEFORMAT 24 Human bax gene retroviral expression vector and its expression in Hela cells Author: CHENG Yu-feng WANG Lai-Cheng Chen Xiujun Abstract [Objective] To construct with the EGFP reporter gene expression of bax gene retroviral vector, the establishment of high expression of bax gene in Hela cells. [Methods] RTPCR method from the Hela cell clones bax gene, according to principles of genetic engineering by PCR, restriction enzyme digestion and other methods to build bax gene and reporter gene EGFP double gene retroviral expression vector pLBaxSNIRES2EGFP, transfected Hela cells, fluorescence microscope and Western Blotting Detection of bax gene expression. [Results] successfully constructed with the EGFP of the bax gene retroviral expression vector and transferred to Hela cells. [Conclusion] successfully established a high expression of bax gene in Hela cell line, in order to explore the bax gene and radiation sensitivity of the relationship between the foundation. 【Key Words】 bax gene retroviral expression vector IRES sequence of vectors Recombinant Human bax Gene Retroviral Vector Construction and Its Expression in Hela Cells Abstrac: [Purpose] To construct the human bax gene retroviral expressive vector with EGFP gene and obtain a Hela strain overexpressing bax gene by transfection. [Methods] Bax gene was cloned from Hela cell with RTPCR. The retroviral vector carrying both bax and EGFP gene was constructed using PCR and enzyme digestion etc, then this new vector pLBaxSNIRES2EGFP was used to transfect Hela cell to establish a new Hela strain, and the expression of the exogenous bax gene was verified by fluorescence microscope and Western Blotting. [Results] The recombinant retroviral vector with exogenous bax and EGFP gene was constructed, and the recombinant vector was transfected into Hela cell. [Conclusion] The successful establishment of Hela cell strain that overexpresses exogenous bax gene provides the