[Construction of recombinant adenovirus vector carrying cell cycle controlling gene-p14ARF].
OBJECTIVE
The recombinant adenovirus vector carrying p14ARF gene was constructed for using in the interference therapy in signal transduction of laryngeal squamous cell carcinoma.


METHODS
The total cDNA fragment of p14ARF was cloned into the shuttle plasmid pAdTrack-CMV, with the resultant plasmid and the backbone plasmid pAdEasy-1, the homologous recombination took place in the E.Coli BJ5183 and the recombinant adenoviral plasmid was generated. The adenoviruses were packaged and amplified in the 293 cells. Then the viral titer was checked by GFP.


RESULTS
The recombinant adenovirus vector carrying p14ARF was constructed successfully. The viral titer was 2.3 x 10(9).


CONCLUSIONS
The recombinant adenovirus vector could introduce p14ARF gene into the laryngeal squamous cell carcinoma line or tumor tissue effectively, which would provide experimental basis for the mechanisms and further study of the interference therapy in signal transduction of laryngeal squamous cell carcinoma.
Xian
Jun-Ming
JM
Dept. of Human Anatomy, Institute of Preclinical Medicine and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Zhou
Guang-Yao
GY
Liang
Chuan-Yu
CY
Liu
Shi-Xi
SX
D000256:Adenoviridae; D002453:Cell Cycle; D005822:Genetic Vectors; D006801:Humans; D010957:Plasmids; D011995:Recombination, Genetic; D025803:Tumor Suppressor Protein p14ARF
D025803:Tumor Suppressor Protein p14ARF