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

نویسندگان

چکیده مقاله:

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 toxin B subunit (CTB) has been known as a mucosal adjuvant for vaccines and genetic fusions of CTB with several hetroantigens such as stxB and can increase humoral and mucosal immunity response.Materials and Methods: In this study, after primer designing, the ctxB and stxB genes were amplified by PCR and cloned into the pGEM-T vector. The stxB gene with a nonfurin linker was fused to the ctB gene in the pGEM vector via the restriction enzyme method and thereafter the fused genes of ctB-stxB were subcloned in the pET28a(+) as an expression vector. The expressed chimeric protein was induced with IPTG and evaluated via the SDS.PAGE and Western blot techniques. Result: The pET28a (+)/ctxB-stxB expression vector was confirmed by endonuclease digestion, PCR, and sequence analysis. The CTB-STB fusion protein was confirmed by the SDS-PAGE and Western-blot. Conclusion: The CTB-STB recombinant protein can be used as a new and desirable mucosal vaccine for Shigella Dysenteriae type I.

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

Fusion of CtxB with StxB, Cloning and Expression of in Esherichia coli: A challenge for Improvement of Immune Response Against StxB

       Cholera toxin B subunit (CtxB) is a homopantameric, nontoxic subunit of cholera toxin that is responsible for its binding to the cell and has been known as a mucosal adjuvant for vaccines that could increase homoral and mocusal immunity response. In this work, the CtxB gene was fused to the StxB gene from Shigella dysenteriae type I a vaccine antigen candidate against t...

متن کامل

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 Clostridium perfringens type D and B epsilon and beta toxin genes and it’s cloning in E. coli

Designing and producing a proper fusion construction is the most important problem of producing large quantities of a properly folded functional protein. This construction should have all necessary components of a real gene. A good designed fusion gene construction could be cloned into a good and suitable host. Clostridium perfringens is an important pathogen of humans and livestock and produce...

متن کامل

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

Molecular Cloning of Clostridium Perfringens type B Vaccine Strain Beta Toxin Gene in E. coli

Clostridium perfringens is a gram-positive, obligate anaerobic bacterium, which is widely distributed in the environment. C. perfringens is subdivided to 5 groups (types A to E), based on its four major toxin (alpha, beta, epsilon and iota). C. perfringens type B beta toxin causes inflammation and bloody necrotic enteritis. Type B related enterotoxaemia is a major problem of veterinary sciences...

متن کامل

منابع من

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

ذخیره در منابع من قبلا به منابع من ذحیره شده

{@ msg_add @}


عنوان ژورنال

دوره 2  شماره 3

صفحات  135- 141

تاریخ انتشار 2012-12

با دنبال کردن یک ژورنال هنگامی که شماره جدید این ژورنال منتشر می شود به شما از طریق ایمیل اطلاع داده می شود.

کلمات کلیدی

کلمات کلیدی برای این مقاله ارائه نشده است

میزبانی شده توسط پلتفرم ابری doprax.com

copyright © 2015-2023