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Updated: Jun 7, 2026
An Efficient Method for Adenovirus Production
Published on: June 10, 2021
[Construction and identification of recombinant adenovirus vector pAdEasy-GFP-GITRL]
1Department of Immunology, School of Medical Science and Laboratory Medicine, Jiangsu University, Zhengjiang 212013, China.
Aim:
To construct recombinant adenovirus vector pAdEasy-GFP-GITRL and detect the viral titer.
Methods:
GITRL gene was obtained by double digestion using Bgl II and Sal I, and cloned into the baculovirus transfer vector(pAdtrack-CMV), then the recombinant adenovirus vector (pAdtrack-CMV-GITRL) was digested by restrictive endoenzyme Pme I. The linear recombinant adenorirus vector and pAdEasy-1 were cotransfected into HEK293 cells by co-precipitate of calcium phosphate. Recombinant adenovirus was packaged and purified in HEK293A cells.
Results:
Recombinant adenovirus vector pAdEasy-GFP-GITRL was constructed successfully and high titer of recombinant adenovirus was obtained (2.0 x 10⁹ pfu/mL). Western blotting analysis also revealed the expression of GITRL by recombinant adenovirus vector.
Conclusion:
The construction of recombinant adenovirus vector pAdEasy-GFP-GITRL and recombinant adenovirus will facilitate the potential GITRL gene therapy.
