[Construction of recombinant strain Brevibacillus choshinensis for chimeric borrelia Dbpa antigen production.]

The aim of this work was creation of recombinant chimeric protein using TAKARA expression system Brevibacillus choshinensis with fused gene dbpAAG, which include the parts of dbpAA and dbpAG genes coding the major antigenic determinants of decorinbinding proteins А (DbpA) from two species of borreliosis agents - Borrelia afzelii and Borrelia gаrinii. Such plasmid should be able to support the synthesis of recombinant chimeric polypeptide consisting immunogenic domains of DbpA Borrelia afzelii and Borrelia gаrinii in the stable and soluble forms, that important for effective using in Lyme diseases serodiagnosis. We chose the TAKARA expression system based on the strain Brevibacillus choshinensis and plasmid pNCMO2. It give us possibilities to obtain the scale quantity of the secreted soluble target proteins with native conformation in particular with conserve antigenic determinants. As results, the plasmid pNCMO2 with a fusion gene dbpAAG was constructed. Recombinante plasmide DNA pNCMO2/dbpAAG was used for Brevibacillus choshinensis trasformation. We were able to show that during cultivation in a liquid medium recombinant cells of В. choshinensis/pNCMO2/dbpAAG produced secreted chimeric 30кD protein with high immunoreactivity to Lyme borreliosis patient's serum.