Fusion of CtxB with StxB, Cloning and Expression of in Esherichia coli: A challenge for Improvement of Immune Response Against StxB Cloning and coexpression of ctB-stxB fusion in E. coli
Iranian Journal of Pharmaceutical Sciences,
Vol. 7 No. 3 (2011),
1 July 2011
,
Page 185-190
https://doi.org/10.22037/ijps.v7.41320
Abstract
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 this pathogen, by a nonfurin linker then ligated with pGEM vector and subcloned in the pET28a(+) as an expression vector . The CtxB-StxB fusion protein was expressed in Escherichia coli, and purified by a Ni-NTA resin column, then detected molecular weight and immunogenicity by SDS-PAGE and Western-blot. StxB has low molecular weight, so immune response against it is low, while CtxB is a potent mucosal adjuvant. In this method, the CtxB-StxB fusion protein was expressed in Escherichia coli in order to use its natural adjuvanticity of the CtxB which will enhance immune response against StxB, as well as, it will produce immune response against both shiga and
cholera toxins.
- Adjuvant
- Expression;
- CtxB,
- Shigella toxin B subunit (StxB);
- Shigella dysenteriae
How to Cite
References
[2] Niyogi SK. Shigellosis. J Microbiol 2005;133-43.
[3] Pina DG, Johannes L. Cholera and Shiga toxin Bsubunits: thermodynamic and structural considerations for function and biomedical applications. Toxicon 2005; 45: 389-93.
[4] Arêas AP, Oliveira ML, Miyaji EN, Leite LC, Aires KA, Dias WO, Ho PL., Expression and characterization of cholera toxin B-pneumococcal surface adhesin A fusion protein in Escherichia coli: ability of CTB-PsaA to induce humoral immune response in mice. Biochem Biophys Res Commun 2004; 321: 192-6.
[5] Odumosu O, Nicholas D, Yano H, Langridge W.AB toxins: A paradigm switch from deadly to desirable. Toxins 2010; 2: 1612-45.
[6] Sambrook J, Fritsch EF, Maniatis T. Molecular cloning: A laboratory manual, 2nd ed. New York:Cold Spring Harbor Laboratory Press, 1989.
[7] Sur D, Ramamurthy T, Deen J, Bhattacharya SK.Shigellosis: challenges and management issues.Indian J Med Res 2004; 454-62.
[8] Schaetti C. Vaccines for enteric diseases: update on recent developments. Expert Rev Vaccines. 2009;8:1653-5.
[9] Zhu C, Yu J, Yang Z, Davis K, Rios H, Wang B, Glenn G, Boedeker EC. Protection against Shiga toxin-producing Escherichia coli infection by transcutaneous immunization with Shiga toxin subunit B. Clin Vaccine Immunol 2008; 15: 359-66.
[10] Zy H, Li MF, Zhang WJ, Wu XF. Cloning of the ctxB Gene of Vibrio cholerae and its expression in E. coli. Vaccine 2003; 32: 149-52.
[11] Holmgren J, Lonnroth I, Mansson J, Svennerholm L. Interaction of cholera toxin and membrane GM1 ganglioside of small intestine. Proc Natl Acad Sci 1975; 72: 2520-44.
[12] Sanchez J, Holmgren J. Cholera toxin structure, gene regulation and pathophysiological and immunological aspects. Cell Mol Life Sci 2008; 65: 1347-60.
[13] Zhang RG, Scott DL, Westbrook ML, Nance S, Spangler BD, Shipley GG, Westbroock EM. The three-dimensional crystal structure of cholera toxin. J Mol Biol 1995; 251: 563-73.
[14] Maddaloni M, Staats HF. Mucosal vaccine targeting improves onset of mucosal and systemic immunity to botulinum neurotoxin. J Immunol 2006; 177: 5524-32.
[15] Czerkinsky C, Sun JB, Lebens M, Li BL, Rask C, Lindblad M, Holmgren J. Cholera toxin B subunit as transmucosal carrier-delivery and immunomodulating system for induction of anti infectious and anti pathological immunity. Ann NY Acad Sci 1996; 778: 185-93.
[16] Mizuno D, Ide-Kurihara M. Modified pulmonary surfactant is a potent adjuvant that stimulates the mucosal IgA production in response to the Influenza virus antigen. J Immunol 2006; 176:1122-30.
[17] Song H, Zhou L, Fang W, Li Y, Wang X, Fang H, Li X, Wu M, Qiu B. High-level expression of codon optimized foot-andmouth disease virus complex epitopes and cholera toxin B subunit chimera in Hansenula polymorpha. Biochem Biophys Res Commun 2004; 235-9.
- Abstract Viewed: 320 times
- IJPS_Volume 7_Issue 3_Pages 185-190 Downloaded: 250 times