Search published articles


Showing 2 results for Cholera Toxin
Designing, Optimization and Construction of Myelin Basic Protein Coding Sequence Binding to the Immunogenic Subunit of Cholera Toxin
N Hashemi, Y Yazdani,
Volume 8, Issue 3 (8-2014)
Abstract
Abstract Background and Objectives: Multiple sclerosis (MS) is a chronic inflammatory autoimmune disease. Mucosal feeding of myelin basic protein binding to the cholera toxin B subunit can reduce the intensity of the immune response in MS patients. Expression system, the domain composition of the fusion protein, accessibility of two domains, codon adaptation index (CAI) and GC contents are very important for the large scale production of fusion protein. Material and Methods: we used DNA2, PSIPRED and ProtParam softwares for designing the best form to produce fusion protein. Moreover, the correct open reading frame of myelin basic protein was also considered. First the coding sequence was verified and then synthesized. For confirmation of the recombinant vector, PCR test was carried out using T7 primers. Finally it was inserted into the cloning site of pET28 expression vector. Results: After coding optimization, the CAI rate was increased from 64 % to 80% and GC content from 41 % to 49%. The presence of a band near 700bp resulted from PCR amplification test demonstrates the correct cloning of recombinant vectors in the cloning site of pET28 expression vector. Conclusion: According to software and experimental analysis, the designed sequence probably in the best form could be used for production of recombinant protein. Keywords: Multiple Sclerosis, Cholera Toxin, Myelin Basic Protein
Cloning, expression, and purification of recombinant Enterotoxin B and cholera toxin B fusion protein in Lactobacillus plantarum
Fatemeh Maghsood Ahmadi , Arash Mahboubi , Farzaneh Hosseini , Davoud Esmaeili , Bahareh Hajikhani ,
Volume 19, Issue 5 (9-2025)
Abstract
Background: Lactic acid bacteria (LAB) are promising platforms for mucosal vaccine development. Staphylococcus aureus enterotoxin B (SEB), a potent superantigen, is associated with food poisoning and toxic shock syndrome. Similarly, cholera toxin is the primary to widespread virulence factor in Vibrio cholerae infections, with its B subunit (CTB) serving as a well-established immune adjuvant that enhances antigen-specific responses in recombinant vaccines. This study aimed to engineer recombinant Lactobacillus plantarum as a dual-purpose vaccine candidate targeting V. cholerae and S. aureus by expressing CTB and SEB antigens.
Methods: A modified gene sequence encoding SEB (Lacking superantigenic activity) and CTB was successfully designed, synthesized, and cloned for secretory expression in L. plantarum. The resulting recombinant protein, tagged with His, was purified using Ni-NTA agarose ion-exchange chromatography and confirmed with Western blot analysis.
Results: Enzyme digestion and PCR analysis confirmed successful cloning of the SEB-CTB fusion gene into the pBlueScript II SK (+) and pNZ7021 expression vectors, as evidenced by the expected band on agarose gel. SDS-PAGE revealed a ~49 kDa protein band, indicating expression of the recombinant rSEB-CTB protein, which was further validated by Western blot using an anti-His tag antibody.
Conclusion: The construct LP-pNZ7021–SP-seb-ctxB may be a promising candidate for recombinant vaccine development targeting V. cholerae (Cholera toxin-producing) and S. aureus (SEB-producing), providing dual protection against both pathogens.
 

Page 1 from 1