The processes of hemostasis and thrombolysis are elegantly regulated in order to ensure normal functions of vascular system. A search for new plasminogen activators as thrombolytic agents has been carried out for the purpose of clinical applications to modulate thrombolytic processes. In the current work, several strains and clinical isolates of Staphylococcus aureus were screened for the fibrinolytic activity. The DNA sequences of staphylokinase gene in the strains expressing 15 kDa protein with staphylokinase activity were determined and subsequently compared with three known staphylokinase gene sequences. From the sequence comparison a new variant of staphylokinase gene has been identified in ATCC 29213 strain. The gene product needs to be further characterized and tested for the therapeutic potential.