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

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 cur...

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Autores principales: Golshani, Maryam, Vaeznia, Nafise, Sahmani, Mehdi, Bouzari, Saeid
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Korean Vaccine Society 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4742603/
https://www.ncbi.nlm.nih.gov/pubmed/26866027
http://dx.doi.org/10.7774/cevr.2016.5.1.75
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author Golshani, Maryam
Vaeznia, Nafise
Sahmani, Mehdi
Bouzari, Saeid
author_facet Golshani, Maryam
Vaeznia, Nafise
Sahmani, Mehdi
Bouzari, Saeid
author_sort Golshani, Maryam
collection PubMed
description 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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spelling pubmed-47426032016-02-10 In silico analysis of Brucella abortus Omp2b and in vitro expression of SOmp2b Golshani, Maryam Vaeznia, Nafise Sahmani, Mehdi Bouzari, Saeid Clin Exp Vaccine Res Original Article 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. The Korean Vaccine Society 2016-01 2016-01-27 /pmc/articles/PMC4742603/ /pubmed/26866027 http://dx.doi.org/10.7774/cevr.2016.5.1.75 Text en © Korean Vaccine Society. http://creativecommons.org/licenses/by-nc/3.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Article
Golshani, Maryam
Vaeznia, Nafise
Sahmani, Mehdi
Bouzari, Saeid
In silico analysis of Brucella abortus Omp2b and in vitro expression of SOmp2b
title In silico analysis of Brucella abortus Omp2b and in vitro expression of SOmp2b
title_full In silico analysis of Brucella abortus Omp2b and in vitro expression of SOmp2b
title_fullStr In silico analysis of Brucella abortus Omp2b and in vitro expression of SOmp2b
title_full_unstemmed In silico analysis of Brucella abortus Omp2b and in vitro expression of SOmp2b
title_short In silico analysis of Brucella abortus Omp2b and in vitro expression of SOmp2b
title_sort in silico analysis of brucella abortus omp2b and in vitro expression of somp2b
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4742603/
https://www.ncbi.nlm.nih.gov/pubmed/26866027
http://dx.doi.org/10.7774/cevr.2016.5.1.75
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