production of pentameric cholera toxin b subunit in escherichia coli

نویسندگان
چکیده

cholera toxin b subunit (ctb) has been extensively studied as an immunogen, adjuvant, and inducer of oral tolerance in many investigations. production of ctb has been carried out in the bacterial, plant, insect and yeast expression systems. in this study the expression of the ctb containing a 6xhis-tagged was performed by escherichia coli (e.coli) m15. the yield of purified pentameric recombinant ctb was about 1 mg/l. western blot analysis demonstrated that the recombinant ctb was antigenically active. in addition, gm1-ganglioside elisa showed that recombinant ctb binds to gm1-gangelioside receptor, confirming disulfide bond formation and proper folding of the recombinant protein in e.coli. overall, in regard to the vast applications of ctb in medicine, this bacterial expression system will be a fast, cost-effective and simple system for production of pentameric ctb and ctb conjugated proteins.

برای دانلود باید عضویت طلایی داشته باشید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Production of Pentameric Cholera Toxin B Subunit in Escherichia coli

Cholera toxin B subunit (CTB) has been extensively studied as an immunogen, adjuvant, and inducer of oral tolerance in many investigations. Production of CTB has been carried out in the bacterial, plant, insect and yeast expression systems. In this study the expression of the CTB containing a 6XHis-tagged was performed by Escherichia coli (E.coli) M15. The yield of purified pentameric recombina...

متن کامل

Fusion of Cholera toxin B subunit (ctxB) with Shigella dysenteriae type I toxin B subunit (stxB), Cloning and Expression that in E. coli

Background and Objective: Shiga toxin (STx) is the main virulence factor in Shigella Dysenteriae type I and is composed of an enzymatic subunit STxA monomer and a receptor-binding STxB homopentamer. Shigella toxin B subunit (STxB) is a non-toxic homopentameric protein responsible for toxin binding and internalization into target cells by interacting with glycolipid (Gb3). Cholera toxi...

متن کامل

ESCHERICHIA COLI HEAT-LABILE TOXIN B SUBUNIT: CONSTRUCTION AND EVALUATION OF PLASMIDS PROVIDING CONTROLLED HIGH LEVEL PRODUCTION OF THE PROTEIN

With the plasmid DNA from a clinical isolate of enterotoxigenic Escherichia coli (ETEC) H 10407 as template, PCR-mediated cloning of the sequence encoding the heat-labile toxin B subunit (L T -B) has been carried out. Then this sequence was recloned into the pTrc 99A and pET23a expression vectors to give the pJasmids pTRCLTB and pETLTB, respectively. After induction, the former plasmid provides...

متن کامل

Chloramphenicol improved expression of recombinant cholera toxin B subunit in Escherichia coli and its adjuvanticity.

BACKGROUND Cholera toxin B subunit (CTB) was shown to be a potent adjuvant for protein immunogen, especially when inoculated through mucosal route. We aimed to optimize the expression approach for CTB and thereafter to determine the adjuvant effect on DNA vaccine. METHODS Wild type CTB coding gene was amplified and cloned into prokaryotic expression vector pET-30a, and the recombinant CTB was...

متن کامل

Expression of functional pentameric heat-labile enterotoxin B subunit of Escherichia coli in Saccharomyces cerevisiae.

Although the Escherichia coli heat-labile enterotoxin B subunit (LTB) has already been expressed in several different systems, including prokaryotic and eukaryotic organisms, studies regarding the synthesis of LTB into oligomeric structures of pentameric size in the budding yeast Saccharomyces cerevisiae have been limited. Therefore, this study used a functional signal peptide of the amylase 1A...

متن کامل

منابع من

با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید


عنوان ژورنال:
avicenna journal of medical biotechnology

جلد ۴، شماره ۲، صفحات ۸۹-۹۴

میزبانی شده توسط پلتفرم ابری doprax.com

copyright © 2015-2023