[Construction of the recombinant adenovirus vector expressing human CD40 ligand gene].
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.


CONCLUSIONS
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.
Wu
Hong-bing
HB
Dept of Radiology, West China College of Stomatology, Sichuan University, Chengdu, China.
Tian
Ling
L
Wen
Yan-jun
YJ
Liu
Yu-mei
YM
Kan
Bing
B
Du
Xiao-bo
XB
Xu
Jian-rong
JR
Wei
Yu-quan
YQ
D000256:Adenoviridae; D000260:Adenoviruses, Human; D000818:Animals; D023201:CD40 Ligand; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D010957:Plasmids; D016133:Polymerase Chain Reaction
D023201:CD40 Ligand