[Construction of a recombinant adenovirus carrying endostatin gene]

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

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.

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.