Construction of a recombinant adenovirus carrying endostatin gene / 南方医科大学学报

ABSTRACT

<p><b>OBJECTIVE</b>To construct a recombinant adenovirus carrying human endostatin gene with AdEasy system.</p><p><b>METHODS</b>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.</p><p><b>RESULTS AND CONCLUSION</b>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.</p>