Pakistan Journal of Biological Sciences1028-88801812-5735Asian Network for Scientific Information10.3923/pjbs.2007.2146.2151RezaNejadmoghaddam Mohammad Mohammad HosseinModarressi MohammadBabashamsi MahmoodChamankhah 1220071013Streptokinase is a common fibrinolytic drug and included in the World Health Organization (WHO) Model List of Essential Medicines. Comparative clinical trails such as cost-effectiveness suggest that streptokinase can be the drug of choice for thrombolytic therapy. To reach the highest amount of the protein and production of active form of streptokinase in bacteria need to modify and optimize methods. In the present study, chromosomal DNA was extracted from S. equisimilis H46A and used for amplification of streptokinase gene (skc) (mature section: 1245 bp) by cloning into pGEX-4T-2 vector which contains a tac promoter. The cloning results were controlled by PCR, double digestion and sequencing. The expression level of the protein in different strain of E. coli was optimized and reached up to 50% of the total cell protein. The function of the fusion protein as active fibrinolytic protein was confirmed by plasmin hydrolysis of chromogenic peptidyl anilide substrate assay.]]>Avilan, L., A. Yarzabal, C. Jurgensen, M. Bastidas, J. Cruz and J. Puig,1997Escherichia coli]]>3014271430Banerjee, A., Y. Chisti and U. Banerjee,200422287307Caballero, A.R., R. Lottenberg and K.H. Johnston,1999Streptococcus equisimilis.]]>6764786486Christensen, L.R.,1945Streptococcal fibrinolysin.]]>28363383 Erdogan, S., A.Y. Ozer, B. Volkan, B. Caner and H. Bilgili,200613303309Estrada, M.P., L. Hernandez, A. Perez, P. Rodriguez and R. Rubiera et al.,1992E. coli.]]>1011381142Gase, K., T. Ellinger and H. Malke,1995Streptococcus equisimilis H46A.]]>247749758Gase, K., A. Gase, H. Schirmer and H. Malke,1996Streptococcus equisimilis H46A gapC gene encoding a glyceraldehyde-3-phosphate dehydrogenase that also functions as a plasmin (ogen)-binding protein. Purification and biochemical characterization of the protein.]]>2394251Hermentin, P., T. Cuesta-Linker and J. Weisse, K.H. Schmidt, M. Knorst, M. Scheld and M. Thimme,200526933940Kazemi, B., F. Fallahian, N. Seyed, M. Bandepour, A. Shabani and P. Ghadam,20069546548Ko, J.H., D.K. Park, I.C. Kim, S.H. Lee and S.M. Byun,1995Escherichia coli.]]>1710191024Kumar, R. and J. Singh,20042113431358Kulisek, E.S., S.E. Holm and K.H. Johnston,19891777884Lee, S.H., S.T. Jeong, I.C. Kim and S.M. Byun,199741199207Malke, H., B. Roe and J.J. Ferretti,1985Streptococcus equisimilis H46A.]]>34357362Nejadmoqaddam, M.R. and M.H. Modaresi,2005Sambrook, J. and D.W. Russell,2003Sazonova, I.Y., A.K. Houng, S.A. Chowdhry, B.R. Robinson, L. Hedstrom and G.L. Reed,20012761260912613Smith, D.B. and K.S. Johnson,1988Escherichia coli as fusions with glutathione S-transferase.]]>673140Sugimura, K. and N. Higashi,1988Escherichia coli.]]>17036503654World Health Organization,20052005Zhai, P., N. Wakeham, J.A. Loy and X.C. Zhang,200342114120