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Iranian Journal of Basic Medical Sciences، جلد ۱۶، شماره ۹، صفحات ۹۸۵-۹۸۹
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عنوان انگلیسی Expression of Recombinant Streptokinase from Streptococcus Pyogenes and Its Reaction with Infected Human and Murine Sera
چکیده انگلیسی مقاله Objective(s): Streptokinase (SKa) is an antigenic protein which is secreted by Streptococcus pyogenes. Streptokinase induces inflammation by complement activation, which may play a role in post infectious diseases. In the present study, recombinant streptokinase from S. pyogenes was produced and showed that recombinant SKa protein was recognized by infected human sera using Western blot analysis. Materials and Methods: In this study, the ska gene from S. pyogenes was amplified and cloned into pET32a which is a prokaryotic expression vector. pET32a-ska was transformed to Escherichia coli BL21 (DE3) pLysS and gene expression was induced by IPTG. Protein production was improved by modification of composition of the bacterial culture media and altering the induction time by IPTG. The expressed protein was purified by affinity chromatography using the Ni-NTA resin. The integrity of the product was confirmed by Westernblot analysis using infected mice. Serum reactivity of five infected individuals was further analyzed against the recombinant SKa protein. Results: Data indicated that recombinant SKa protein from S. pyogenes can be recognized by patient and mice sera. The concentration of the purified recombinant protein was 3.2 mg/L of initial culture. The highest amount of the expressed protein after addition of IPTG was obtained in a bacterial culture without glucose with the culture optical density of 0.8 (OD600 = 0.8). Conclusion : Present data shows, recombinant SKa protein has same epitopes with natural form of this antigen. Recombinant SKa also seemed to be a promising antigen for the serologic diagnosis of S. pyogenes infections.
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نویسندگان مقاله ندا مولایی | neda molaee
department of cellular and molecular biology, science and research branch, islamic azad university, tehran, iran

سازمان اصلی تایید شده: دانشگاه آزاد اسلامی علوم و تحقیقات (Islamic azad university science and research branch)

حمید ابطحی | hamid abtahi
molecular and medicine research center, arak university of medical sciences, arak, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی اراک (Arak university of medical sciences)

قاسم مسیبی | ghasem mosayebi
molecular and medicine research center, arak university of medical sciences, arak, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی اراک (Arak university of medical sciences)

نشانی اینترنتی http://ijbms.mums.ac.ir/article_1679.html
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زبان مقاله منتشر شده en
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نوع مقاله منتشر شده Original Article
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