[Construction of a recombinant adenovirus carrying endostatin gene]

Ying Ma, Yuan-Li He, Fang Yang
Nan Fang Yi Ke da Xue Xue Bao, Journal of Southern Medical University 2007, 27 (10): 1514-6

OBJECTIVE: To construct a recombinant adenovirus carrying human endostatin gene with AdEasy system.

METHODS: Endostatin gene fragment was amplified from Pshuttle-Endostatin plasmid with PCR and subcloned into the pAdTrack-CMV shuttle vector. The resultant plasimid was cotransduced into E.coli BJ 5183 cells with pAdEasy-1 plasmid for homologous recombination. The linearized recombinant plasmid was subsequently transfected into AAV 293 cells, and the recombinant adenovirus was detected by GFP, PCR and restriction analysis.

RESULTS AND CONCLUSION: The positive clones of the recombinants were verified by restriction analysis and the titer of the virus reached 2.06 x 10(10)pfu/ml, suggesting successful construction of recombinant adenovirus pAd-Endo.

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