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Updated: Jun 4, 2026
An Efficient Method for Adenovirus Production
Published on: June 10, 2021
[Study on construction of chimeric adenovirus vector Ad5/11 carrying human eGFP and endostatin-K5 and its
Dong-Yang Wang1, Shi-Hai Liu, Xing Li
1Laboratory of Gene Therapy, College of Life Sciences, Shaanxi Normal University, Xi'an 710062, China. wangdongyang007@yahoo.com.cn
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.
