Design and Production of a Novel Recombinant Chimeric IL2-Omp31 Antigen against Brucella Infection

Document Type: Original Articles

Authors

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

Abstract

Brucellosis is a zoonotic disease in human and animals. Brucella melitensis is one of the most pathogenic species of Brucella in goat and sheep. Omp31 is an outer membrane protein of Brucella that acts as an immunogenic protein. Cytokines are glycoproteins with low molecular weight that play the role of an immune adjuvant and regulate immune responses. Interleukin-2 is one of the most important cytokines, which are secreted by the white blood cells and involved in T cell immune responses. In the present study, a chimeric Omp31-Interleukin2 recombinant protein was generated by means of genetic engineering techniques. This chimeric coding sequence was amplified by using specific primers and using Splicing Overlap Extension (SOE) PCR technique. The fusion of the two mentioned proteins was accomplished using a rigid linker. The generated chimeric IL2-Omp31 fragment was TA cloned, and then subcloned into pEt22b vector as an expression vector. The chimeric protein was successfully expressed in E. coli BL21 (DE3) and confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and also Western-blotting analysis. Finally, in order to assess the antigenic features of the recombinant chimeric IL2-Opm31 protein, its secondary structure and antigenicity were predicted in silico.

Keywords

Main Subjects


Article Title [French]

Conception et Production d'un Nouvel Antigène Chimique IL2-Omp31 Recombinant Contre l'infection à Brucella

Abstract [French]

La brucellose est une maladie zoonotique chez l'homme et les animaux. Brucella melitensis est l'une des espèces les plus pathogènes de Brucella chez les chèvres et les moutons. Omp31 est une protéine de membrane externe de Brucella qui agit comme une protéine immunogène. Les cytokines sont des glycoprotéines de faible poids moléculaire qui jouent le rôle d'un adjuvant immunitaire et régulent les réponses immunitaires. L'interleukine-2 est l'une des cytokines les plus importantes, sécrétées par les globules blancs et impliquées dans les réponses immunitaires des cellules T. Dans la présente étude, une protéine recombinante chimère Omp31-Interleukine2 a été générée au moyen de techniques de génie génétique. Cette séquence de codage chimérique a été amplifiée en utilisant des amorces spécifiques et en utilisant la technique de PCR par Extension de Chevauchement d'Épissage" (ECE). La fusion des deux protéines mentionnées a été réalisée en utilisant un connecteur rigide. Le fragment chimérique d'IL2-Omp31 généré a été TA cloné, puis sous-cloné dans le vecteur pEt22b en tant que vecteur d'expression. La protéine chimère a été exprimée avec succès dans E. coli BL21 (DE3) et confirmée par électrophorèse sur gel de dodécylsulfate de sodium polyacrylamide (SDS-PAGE) et également par analyse par transfert Western. Enfin, afin d'évaluer les caractéristiques antigéniques de la protéine chimérique IL2-Opm31 recombinante, sa structure secondaire et son antigénicité ont été prédites in silico.

Keywords [French]

  • Brucella melitensis
  • Interleukine-2
  • Omp31
  • Cytokines
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