Cloning and molecular characterization of Omp31 gene from Brucella melitensis Rev 1 strain

Document Type: Short Communication

Authors

Department of Animal Sciences, Faculty of Agriculture, Ferdowsi University of Mashhad, Mashhad, Iran

Abstract

Brucellosis, caused by the genus Brucella bacterium, is a well-known infection among domestic animals. Considering the serious economic and medical consequences of this infection, various preventive efforts have been made through using recombinant vaccines, based on outer membrane protein (OMP) antigens of Brucella species. The objective of the present study was to clone, analyze the sequence, and predict the epitopes of Omp31 gene as a major B. melitensis antigen. The full-length open reading frame (ORF) for this gene was amplified by specific primers and cloned into the pTZ57R/T vector. The gene sequence of B. melitensis Rev 1 strain was submitted to NCBI database. The results of phylogenetic analysis showed that Omp31 is almost similar in different Brucella species. Online prediction software programs were also used to predict B- and T-cell epitopes, secondary and tertiary structures, antigenicity, and enzymatic degradation sites. The bioinformatic tools in the current study were confirmed by the results of three different experimental epitope prediction studies. Bioinformatic analysis identified one T-cell and three B-cell epitopes for Omp31 antigen. Finally, based on the antigenicity and proteosome recognition sites, common B- and T-cell epitopes were predicted for Omp31 (amino acids 191-204). Bioinformatic analysis showed that these regions had proper epitope characterization and could be useful for recombinant vaccine development.

Keywords

Main Subjects


Article Title [French]

Clonage et caractérisation moléculaire du gène Omp31 de la souche Rev 1 de Brucella melitensis

Abstract [French]

La brucellose, une infection causée par des bactéries du genre Brucella, est très répandue chez les animaux domestiques. Compte tenu des conséquences économiques et sanitaires de cette infection, des mesures de prévention variées ont été mises en place, comme notamment l’usage de vaccins recombinants à partir d’antigènes isolés des membranes cellulaires externes des espèces de Brucella. L’objectif de cette étude est le clonage, l’analyse et le séquençage du gène Omp31 ainsi que la prédiction des épitopes de l’antigène majeur de B. melitensis. Le cadre de lecture ouvert de pleine longueur (full-length open reading frame, ORF) du gène Omp31 a été amplifié grâce à l’utilisation d’amorces spécifiques et ensuite cloné dans le vecteur pTZ57R/T. La séquence du gène a été soumise à la base de données NCBI. Les résultats de nos analyses phylogénétiques montrent qu’Omp31 est pratiquement similaire dans toutes les espèces du genre Brucella. Des logiciels de simulation en ligne ont été également utilisés afin de prédire les séquences et l’emplacement des épitopes spécifiques aux cellules B et T, les structures secondaire et tertiaire de l’antigène ainsi que les sites probables de dégradation enzymatiques. Nos analyses bioinformatiques ont permis d’identifier un épitope de cellule T et trois épitopes de cellules B pour l’antigène Omp31. Enfin, à partir de la reconnaissance des sites antigéniques et protéasomiques, des épitopes reconnues communément par les cellules B et T ont été identifiés pour l’antigène Omp31 (acides aminés 191-204). Nos analyses bioinformatiques montrent que des régions spécifiques de l’antigène Omp31 exhibent des propriétés épitopiques et pourrait être utilisées pour le développement futur de vaccin recombinant.

Keywords [French]

  • Brucella melitensis
  • Omp31
  • Analyse bioinformatique

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