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KMID : 1039120160050010075
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Clinical and Experimental Vaccine Research
2016 Volume.5 No. 1 p.75 ~ p.82
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In silico analysis of Brucella abortus Omp2b and in vitro expression of SOmp2b
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Golshan Maryam
Vaeznia Nafise
Sahmani Mehdi
Bouzari Saeid
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Abstract
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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 μg/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.
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KEYWORD
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Brucella, Omp2b, In silico approach, Epitope prediction, Protein expression
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