Clin Exp Vaccine Res.  2016 Jan;5(1):75-82. 10.7774/cevr.2016.5.1.75.

In silico analysis of Brucella abortus Omp2b and in vitro expression of SOmp2b

Affiliations
  • 1Department of Molecular Biology, Pasteur Institute of Iran, Tehran, Iran. saeidbouzari@yahoo.com
  • 2Department of Biotechemistry, Qazvin University of Medical Sciences, Qazvin, Iran.

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.

Keyword

Brucella; Omp2b; In silico approach; Epitope prediction; Protein expression
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