In silico analysis of Omp25 and BLS Brucella melitensis antigens for designing subunit vaccine

Document Type: Original Articles

Authors

Department of Animal Science, Ferdowsi University of Mashhad, Mashhad, Iran

Abstract

Brucellosis is a well-known infection in domestic animals which caused by Brucella bacterium. Due to
serious economic and medical consequences of this disease, various efforts have been made to prevent the
infection through the use of recombinant vaccines based on Brucella outer membrane protein (OMP)
antigens. The objectives of the present study were cloning, sequencing and epitope prediction of Omp25
and BLS genes as two major Brucella melitensis antigens. The full-length open reading frame (ORF) of
Omp25 and BLS genes were amplified and cloned into pTZ57R/T vector. Phylogenetic analysis of
sequenced genes showed that both genes were nearly similar in different Brucella species. Several online
prediction softwares were used to predict B and T-cells epitopes, secondary and tertiary structures,
antigenicity ability and enzymatic degradation sites. Bioinformatic tools used in the current study were
confirmed by the results of three different experimental epitope predictions. Bioinformatic analysis
identified five and two B-cell and two and one T-cell epitopes for Omp25 and BLS antigens, respectively.
Finally, according to the antigenicity ability and proteosomal recognition site common B and T-cell epitope
was predicted for Omp25 (154-162 amino acids) and BLS (37-48 and 119-139 amino acids). Results of this
study might be useful for recombinant vaccine development.

Keywords

Main Subjects


Article Title [French]

Analyse in silico des antigènes Omp25 et BLS de Brucella melitensis pour l’élaboration d’un vaccin de sous-unité

Abstract [French]

La brucellose est une infection qui touche particulièrement les animaux domestiques et qui est causée par les bactéries du genre Brucella. En raison des conséquences économiques et médicales importantes de cette maladie, diverses efforts on été conduits dans l’élaboration de vaccins recombinants à partir de protéines antigéniques de la membrane externe (OMP). L’objectif de cette étude était le séquençage, le clonage et la prédiction des épitopes des gènes « Omp25 et BLS » codant les deux antigènes majeurs de Brucella melitensis. Le cadre de lecture ouvert (ORF) pleine longueur des gènes Omp25 et BLS a été amplifié et cloné dans le vecteur pTZ57R/T. L’analyse phylogénétique des séquences obtenues montrent que les deux gènes sont presque semblables dans les différentes espèces du genre Brucella. Plusieurs logiciels prévisionnels ont été utilisés pour prédire l’emplacement des épitopes des cellules B et T, les structures secondaires et tertiaires, le potentiel antigénique et les sites de clivage enzymatique des antigènes d’intérêt. Les résultats de nos tests bioinformatiques ont été confirmés par 3 méthodes différentes de prédiction expérimentale des épitopes. Ces analyses ont révélé que le nombre d’épitopes de cellules B était de 5 pour Omp25 et 2 pour BLS, alors que seulement 2 et 1 épitopes de cellules T ont été respectivement identifiés pour ces deux antigènes. Enfin, l’étude du potentiel allergénique et du protéasome des antigènes, nous a permis de prédire la présence d’épitopes de cellules B et T communs aux niveaux des acides aminés numéros 154 à 162 pour Omp25 et 37 à 48 ainsi que 119 à 139 pour BLS. Les résultats de ce travail pourraient être utiles dans l’élaboration future d’un vaccin recombinant.

Keywords [French]

  • Brucella melitensis
  • Omp25
  • BLS
  • Analyse bioinformatique
  • Vaccin recombinant

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