Cloning, expression and purification of rSEB-CTB fusion protein in Lactobacillus plantarum

Maghsood Ahmadi, fatemeh; Mahboubi, Arash; Hosseini, Farzaneh; Esmaeili, Davoud; Hajikhani, Bahareh

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Cloning, expression and purification of rSEB-CTB fusion protein in Lactobacillus plantarum

Fatemeh Maghsood Ahmadi¹

, Arash Mahboubi²

, Farzaneh Hosseini³

, Davoud Esmaeili⁴

, Bahareh Hajikhani⁵

1- aDepartment of Microbiology North Tehran Branch, Islamic Azad University, Teheran, Iran
2- bFood Safety Research Center, Department of Pharmaceutics, School of Pharmacy, Shahid Beheshti University of Medical Sciences, 2660, Vali-Asr Ave, Tehran, Iran. , mahboubi@sbmu.ac.ir
3- Department of Microbiology North Tehran Branch, Islamic Azad University, Teheran, Iran.
4- Department of Microbiology and Applied Microbiology Research Center, Systems biology and poisonings institute, Baqiyatallah University of Medical sciences, Tehran, Iran
5- Department of Microbiology, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, IR Iran

Abstract: (19 Views)

ABSTRACT
Background and objectives: Lactic acid bacteria (LAB) are potential candidates for the mucosal vaccine. Staphylococcal enterotoxin B (SEB), as a potent superantigen exotoxin is associated with widespread dietary poisoning and induction of toxic shock syndrome. Also, cholera toxin is the most important virulence factor in Vibrio cholera pathogenicity. CTB, a well-known immune adjuvant, enhances immunity and is mainly used to produce recombinant vaccines as antigen immunization enhancers. This study aimed to produce recombinant Lactobacillus Plantarum as a candidate vaccine against Vibrio cholera producing Cholera toxin and Staphylococcus aureus producing enterotoxin SEB. Methods: A gene sequence encoding SEB, devoid of superantigenic activity, and CTB were successfully designed, synthesized, cloned, and then expressed in a secreted form in the Lactobacillus Plantarum. The recombinant protein containing His-Tag was purified by Ni-NTA Agarose ion-exchange chromatography column. The purified protein was confirmed by Western blotting. Results: The result of this study demonstrated the expression of this recombinant protein in the Lactobacillus Plantarum system by pnz7021 expression vector. The protein electrophoresis showed that the molecular weight of recombinant fusion protein was 52 kDa. Western blot analysis also confirmed the production of recombinant protein. The use of recombinant vaccines has received a great deal of attention today. LP-pnz7021–SP-rseb-ctxB can be used as a suitable candidate in recombinant vaccines against Vibrio cholera producing Cholera toxin and Staphylococcus aureus producing enterotoxin SEB.

Keywords: rSEB, Lactobacillus Plantarum, CTB

Full-Text [PDF 460 kb] (11 Downloads)

Research Article: Research Article | Subject: Microbiology
Received: 2021/05/4 | Accepted: 2021/06/28

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