Production of Mutant Streptokinase Recombinant Protein

authors:

avatar Negar Seyed 1 , avatar Parvaneh Shekari 2 , avatar Mojgan Bandehpour 1 , avatar Zarrin Sharifnia 1 , avatar Kazem Parivar 2 , avatar Bahram Kazemi ORCID 1 , *

Cellular and Molecular Biology Research Center, Shahid Beheshti University, M.C., Tehran, IR Iran
Islamic Azad University, Research and Science Campus, Tehran, IR Iran

how to cite: Seyed N, Shekari P, Bandehpour M, Sharifnia Z, Parivar K, et al. Production of Mutant Streptokinase Recombinant Protein. Arch Clin Infect Dis. 2008;3(4): 179-183. 

Abstract

Background:

Streptokinase (SK) is most widely used for treatment of myocardial infarction, however, it is the most expensive thrombolytic agent. A major drawback to SK use is the widespread presence of antistreptokinase antibodies (Abs). These Abs cause allergic reactions and neutralize streptokinase therapeutic effects.

Materials and methods:

To produce an engineered variant of streptokinase being functional and less antigenic than the native molecule, we cloned and expressed streptokinase mutant gene lacking the C terminal 42 amino acids. Recombinant protein was confirmed by western blot analysis with anti T7 monoclonal antibodies.

Results:

pGEMEX-1 expression vector contains T7 gene 10 protein as fusion protein immediately down stream of T7 promoter and before multiple cloning site, streptokinase mutant gene was cloned after fusion protein.

Conclusion:

We cloned and expressed mutant streptokinase gene, lacking the C-terminal 42 amino acids. If mut-C42 activity was less affected by neutralizing antibodies compared with native streptokinase, this engineered variant could be a preferred alternative to native streptokinase for thrombolytic therapy.

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