[Streptokinase: extraction, cloning and overexpression of recombinant protein with simplified purification technique]

ABSTRACT

Streptokinase has been widely prescribed as a fibrinolytic agent for myocardial infarction. Simple structural characteristics of this protein have provided techniques for production of different recombinant types of this protein. The present study was designed to prepare equisimilisH46A subtype of streptokinase in our country. Having extracted the DNA, the streptokinase gene was replicated and cloned in pGEX-4T-2. The recombinant product was transformed in BL21 [DE3] plysS'. Then the recombinant streptokinase expression and performance was assessed by lasic densitometry and special test for S2251 substrate. Use of a restriction enzyme for both sides of a gene may facilitate its cloning in different expressive carriers. Expression rate of recombinant protein [45%] confirmed successful cloning. Use of pGEX-4T-2 carrier was not only associated with active recombinant streptokinase production, added GST to its amine ending that could facilitate purification process