[Home ] [Archive]   [ فارسی ]  
:: Main :: About :: Current Issue :: Archive :: Submit :: Contact Us ::
:: Volume 2, Issue 3 (Autumn 2012) ::
J Fasa Univ Med Sci 2012, 2(3): 135-141 Back to browse issues page
Fusion of Cholera toxin B subunit (ctxB) with Shigella dysenteriae type I toxin B subunit (stxB), Cloning and Expression that in E. coli
Hamid Madanchi 1, Hossein Honari, Sadegh Safaei, Ali Sayadmanesh
1- , hamidmadanchi@yahoo.com
Abstract:   (17025 Views)
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.
Keywords: Adjuvant; CTB; Nonfurin linker; StxB; Shigella Dysenteriae type I
Full-Text [PDF 1631 kb]   (3075 Downloads)    
Type of Study: Research | Subject: Microbiology
Received: 2013/01/31 | Accepted: 2013/09/14 | Published: 2013/09/14
Send email to the article author

Add your comments about this article
Your username or Email:

CAPTCHA


XML   Persian Abstract   Print


Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:

Madanchi H, Honari H, Safaei S, Sayadmanesh A. Fusion of Cholera toxin B subunit (ctxB) with Shigella dysenteriae type I toxin B subunit (stxB), Cloning and Expression that in E. coli. J Fasa Univ Med Sci. 2012; 2 (3) :135-141
URL: http://journal.fums.ac.ir/article-1-26-en.html


Volume 2, Issue 3 (Autumn 2012) Back to browse issues page
Journal of Fasa University of Medical Sciences
Persian site map - English site map - Created in 0.06 seconds with 32 queries by YEKTAWEB 4006