Objective To sequence and express tissue type plasminogen activator (tPA) cDNA of rhesus monkey. Method\ The cDNA was obtained by RT PCR from the lymphocyte of rhesus monkey, of which the primers were designed according to human tPA cDNA sequence. The amplified fragment was then cloned into T vector. Results\ The result of nucleotide sequencing showed the coding region of rhesus monkey tPA to be 96% nucleotides and 97.5% amino acids identity with human tPA. In the subsequent experiment, the rhesus monkey tPA cDNA was cloned into eukaryotic expression vector, and tPA activity of 50 U/ml can be detected in the medium at the second day (48hours) after transfection of CHO cells. Conclusion\ The coding region cDNA sequence of rhesus monkey and its expression in mammalian cells are reported. It could be useful for the further study of tPA biological characters.