Chinese Journal of Cellular and Molecular Immunology. 2011, (02) Core

Study on construction of chimeric adenovirus vector Ad5/11 carrying human eGFP and endostatin-K5 and its experimental investigation in vitroChinese Full Text

WANG Dong-yang,LIU Shi-hai,LI Xing,ZHAO Jun-li,CHEN Hao,FENG Fei-xue,WANG Li-xian,XIA Hai-bin* Laboratory of Gene Therapy,College of Life Sciences,Shaanxi Normal University,Xi’an 710062, China

Abstract: AIM:To construct chimeric adenoviral vector Ad5/11 carrying reporter gene eGFP and human endostatin-K5.METHODS:Chimeric adenoviral backbone vector expressing eGFP was generated by overlap PCR and homologous recombination in E.coli BJ5183.Then chimeric adenoviral vector Ad5/11-E1-CMV-endo-K5/E3-CMV-eGFP carrying eGFP and human endostatin-K5 was constructed by co-transfecting Pac I linearized chimeric adenoviral backbone and adenoviral E1 shuttle vector expressing human endostatin-K5 into HEK 293 cells.The expression of eGFP was observed under fluorescent microscope.The expression of human endostain-K5 in U87MG cells infected by chimeric adenoviral vector was detected by RT-PCR.The infection efficiency between chimeric adenovirus and unmodified control adenovirus for human glioblastoma cell line A172 and breast cancer cell line MDA-MB-231 in vitro was evaluated by the comparison of the expression of eGFP.RESULTS:Chimeric adenovirus Ad5/11-E1-CMV-endo-K5/E3-CMV-eGFP could successfully express eGFP and endostatin-K5.Chimeric adenoviral vector significantly enhances the infection efficiency for human glioblastoma cell line A172 and breast cancer cell line MDA-MB-231 compared with unmodified adenoviral vector Ad5 E1-CMV-eGFP.CONCLUSION:Chimeric adenoviral vector Ad5/11-E1-CMV-endo-K5/E3-CMV-eGFP can significantly improve the infection efficiency for human glioblastoma cell line A172 and breast cancer cell line MDA-MB-231.

Keywords：

endostatin-K5; adenoviral vector; tumor; gene therapy;

DOI：
10.13423/j.cnki.cjcmi.005961
Series：
Subject：
Classification Code：
R73-3

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Study on construction of chimeric adenovirus vector Ad5/11 carrying human eGFP and endostatin-K5 and its experimental investigation in vitroChinese Full Text

WANG Dong-yang,LIU Shi-hai,LI Xing,ZHAO Jun-li,CHEN Hao,FENG Fei-xue,WANG Li-xian,XIA Hai-bin* Laboratory of Gene Therapy,College of Life Sciences,Shaanxi Normal University,Xi’an 710062, China

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