Aim To construct the recombinant adeno-associated virus vector(rAAV) expressing the human tissue kallikrein(HK) gene and detect the expression of interested gene in human umbilical vein endothelial cell(hUVEC) which were infected with different titer of rAAV.The feasibility of gene therapy for hypertension by rAAV mediated human tissue kallikrein gene transfection was discussed. Methods The HK gene was directionally cloned into the pAAV-MCS,and co-transfected AAV-293 cell with other two plasmids(the pAAV-RC,and pHelper) by lipofectamine.The recombinant AAV particles were harvested and the viral titer was measured.Then hUVEC were infected with different titer of rAAV particles.72 hours later,the expression of human tissue kallikrein gene was detected by RT-PCR and ELISA. Results The AAV expressing system of human tissue kallikrein gene was successfully constructed with a titer of 6.2×10~(10) particles/L measured with In Situ β-Galactosidase Staining Kit.Compared with control group,the expression of HK gene in groups infected with rAAV at different titer of 1×10~9,1×10~8,1×10~7 increased significantly(p<0.05),especially in the group of 1×10~9(p<0.001). Conclusions When co-transfecting AAV-293 cell with three plasmids(the recombinant pAAV expression plasmid,pAAV-RC,and pHelper),the titer of rAAV particles can reach >10~(10) particle/L stably.The interested gene can be expressed significantly and stably when using recombinant AAV viral to infect the cultured mammal cell.