In order to study the co-expression effect of antisense thrombin receptor (ATR) and p21 genes on preventing restenosis, an adeno-associated virus(AAV) vector carrying the two genes with different promoters was constructed. Simultaneously, rAAV vectors containing ATR or/and p21 gene were constructed respectively. BHK-21 cell lines carried different plasmids were established by lipofectamine transfection and G418 selection. During the process of cell clones selected, double gene showed stronger inhibition to cell proliferation than single gene. The speed of clones formation decreased meanwhile the morphological alteration of cells occurred. Southern blot confirmed that expression vectors containing ATR or/and p21 gene had been steadily integrated into BHK-21 cells. The antisense position of TR gene has been maintained. Semi-quantitative RT-PCR showed that the expression of TR decreased and the expression of p21 increased. Co-expression of double gene was obtained. rAAV virions were packaged through recombinant herpes simplex virus (rHSV-rc/ΔU12) infecting BHK-21 cell lines carried various vectors. The titre of rAAV (particles/ml) determined by dot blot analysis. The particles of ATR and p21 in rAAV/AP double gene vector were 1.02×10¹³ particles/ml and 1.08×10¹³ particles/ml respectively. The particles of single gene rAAV /ATR or rAAV /p21 were 6.54×10¹² particles/ml or 1.06×10¹³ particles/ml.