Human plasminogen activator inhibitor 1 (PAI-1) gene was recombined into a replication-defective adenovirus vector AdCMVHSgD, co-transfected with plasmid pJM17 into human embryo kidney cell line 293 cells. Recombinant virus was produced without use of agarose overlay, identified by PCR analysis and used to infect B16 (F10) cells. The inhibition of plasminogen activator (PA) activity of PAI-1 was revealed in the elute and supernatant of infected B16 cells by regular and reverse agarose-fibrin autography.