In silico analysis of Brucella abortus Omp2b and in vitro expression of SOmp2b.
- Author:
Maryam GOLSHANI
1
;
Nafise VAEZNIA
;
Mehdi SAHMANI
;
Saeid BOUZARI
Author Information
- Publication Type:In Vitro ; Original Article
- Keywords: Brucella; Omp2b; In silico approach; Epitope prediction; Protein expression
- MeSH: Brucella abortus*; Brucella*; Brucellosis; Clone Cells; Cloning, Organism; Computational Biology; Computer Simulation*; Epitopes, B-Lymphocyte; Escherichia coli; Humans; Membrane Proteins; T-Lymphocytes
- From:Clinical and Experimental Vaccine Research 2016;5(1):75-82
- CountryRepublic of Korea
- Language:English
- Abstract: PURPOSE: At present, there is no vaccine available for the prevention of human brucellosis. Brucella outer membrane protein 2b (Omp2b) is a 36 kD porin existed in common Brucella pathogens and it is considered as priority antigen for designing a new subunit vaccine. MATERIALS AND METHODS: In the current study, we aimed to predict and analyze the secondary and tertiary structures of the Brucella abortus Omp2b protein, and to predict T-cell and B-cell epitopes with the help of bioinformatics tools. Subsequently, cloning and expression of the short form of Omp2b (SOmp2b) was performed using pET28a expression vector and Escherichia coli BL21 host, respectively. The recombinant SOmp2b (rSOmp2b) was purified with Ni-NTA column. RESULTS: The recombinant protein was successfully expressed in E. coli host and purified under denaturation conditions. The yield of the purified rSOmp2b was estimated by Bradford method and found to be 220 microg/mL of the culture. CONCLUSION: Our results indicate that Omp2b protein has a potential to induce both B-cell- and T-cell-mediated immune responses and it can be evaluated as a new subunit vaccine candidate against brucellosis.
