Preparation of the antiangiogenic peptide alphastatin recombinant adeno-associated virus

Objective To construct and produce the vector of antiangiogenic peptide alphastatin (Al) recombinant adeno-associated virus (rAAV).Methods By inserting the resulting gene of Al into the EcoR I and BamH I sites of vector plasmid pSSHG- cytomegalovirus (CMV), the recombinant plasmid vector pSSHG-CMV/NT4-Al was constructed. The 293 cells were co-transfected with the recombinant plasmid, packaging plasmid pAAV/Ad and helper adenovirus plasmid pFG140 instead of wild type adenovirus. The rAAV-A1 viral stock was purified by chloroform and polyethylene glycol precipitation and titrated by dot blot.Results The titer of rAAV-A1 vector was approximately 2×10 12 particles/ml.Conclusion The viral stock of rAAV-Al is successfully prepared, which can lay a basis for the experimental studies of gene therapy of tumors.