Related Experiment Video
Updated: Aug 13, 2026
Adenoviral Transduction of Naive CD4 T Cells to Study Treg Differentiation
Published on: August 13, 2013
[Construction of the recombinant adenovirus vector expressing human CD40 ligand gene]
Hong-bing Wu1, Ling Tian, Yan-jun Wen
1Dept of Radiology, West China College of Stomatology, Sichuan University, Chengdu, China.
Objective:
To construct a recombinant adenovirus vector expressing hCD40L gene and explore it in the use of anti-tumor gene therapy.
Methods:
1,900 bp gene fragment was obtained form plasmid pORF-hCD40L by Xho I/Swa I cutting and then cloned directionally into the pShuttle plasmid, finally, the resultant plasmid was digested by restriction endonnuclease PmeI and subsequently cotransformtion into BJ5183 cells with the adenoviral backbone pAdEasy-1 to obtain the homologous recombinant and then the recombinant was packaged in the 293 cells. Some methods such as PCR and endonulease digestion were employed to identify the recombinant adenovirus.
Results:
The evidences of endonulease digestion and PCR analysis confirmed that recombinant hCD40L gene was correctly inserted into adenovirus vector.
Conclusion:
The adenoviral vector which expressed hCD40L gene was constructed. It provides an experimental basis for studies on it expression in the mammalian cells and in tumor gene therapy.
