Development and immunereactivity evaluation of a chimeric recombinant protein encoding <i>Brucella</i> antigen: <i>In silico </i> to <i>in vitro</i>

30-36Brucellosis is an important health problem in developing countries and no vaccine is available for the prevention of infection in humans. Because of clinically infectious disease and its economic consequences in human and animals, designing a proper vaccine against Brucella is desirable. In the present study, we evaluated the immune responses induced by a designed recombinant chimera protein and investigated the immunogenic potential of some immune reactive antigens of Brucella. Three immune dominant antigens of Brucella including trigger factor (TF), Omp31 and Bp26 (have been characterized as potential immunogenic and protective antigens) were fused together by EAAAK linkers to produce a chimera. Recombinant chimeric protein was synthesized, cloned and expressed in Escherichia coli BL21 (structure were designed in silico). The purification of recombinant protein was performed by using Ni-NTA agarose, and anti-His antibody was used for confirmation (Western blot). The recombinant chimeric protein could be a new potential antigen candidate for the development of a subunit vaccine against Brucella. These results demonstrate the role of the Bioinformatics in vaccine design, assisted by experimental procedures