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Updated: Jun 30, 2026
An Efficient Method for Adenovirus Production
Published on: June 10, 2021
[Construction of a recombinant adenovirus expression vector for human renal tumor- associated antigen G250 gene with
1Department of Urology, Nanfang Hospital, Southern Medical University, Guangzhou, China. qihuan@fimmu.com
Objective:
To construct a adevoviral vector harboring human renal tumor-associated antigen G250 gene for transfecting the dendritic cells (DCs) and treating renal tumors.
Methods:
The G250 genes were cloned into the shuttle plasmid pDC316 to construct pDC316-G250, which was cotransfected with the rescue plasmid pBHGlox(delta)E1,3Cre into 293 cells to obtain the recombinant adenovirus Ad/G250. The inserted gene and its expression were identified by RT-PCR and fluorescence-activated cell sorting (FACS) after recombinant adenovirus transfection of the DCs. The recombinant adenoviral vector was purified by CsCl banding and titrated by TCID50.
Results And Conclusion:
The recombinant adenoviral vector of G250 gene was successfully constructed and high titer of the recombinant adenoviruse was obtained. G250 mRNA and protein expressions were identified in Ad/G250-transfected DCs. The titer of the virus stocks reached 5.6x10(9) IU/ml.