Cloning and expression of a chimera containing ROP2 of Neospora caninum fused with OprI lipoprotein from Pseudomonas aeruginosa / Clonagem e expressão de quimera contendo a proteína ROP2 de Neospora caninum fusionada a lipoproteína OprI de Pseudomonas aeruginosa

Abstract

Neospora caninum is the etiologic agent of neosporosis, is one of the main responsible for abortion in cattle herds, causing economic losses to Cattle-raising. Vaccination of cattle would be an important alternative; however, the lack of effective vaccines prevents the application of this control method. The proteins present in rhoptries (ROPs), due to their importance in cell infection and their antigenic and immunogenic characteristics, became excellent candidates for vaccine antigens. The bacterial lipoproteins as an Oprl from Pseudomonas aeruginosa have received particular attention as an adjuvant carrier molecule. The aims of this study were to clone and expressing a chimera containing NcROP2 fused with OprI lipoprotein from P. aeruginosa for the future development of a recombinant vaccine against N. caninum. We cloned and expressed, in Escherichia coli Rosetta (DE3) pLysS, the region of NcROP2 described between amino acids 191 and 359, fused OprI producing the chimera rROP2/OprI, showed an expected size of ~50 kDa. and characterized its antigenicity. The protein was purified and characterized by Western blot with anti-histidine monoclonal antibodies and their antigenicity recognized by sera from animals naturally infected by N. caninum. The chimera rROP2/OprI was recognized by antibodies anti-N. caninum reviling common antigenic determinants of the recombinant protein and its native form, suggesting its use for developing a recombinant vaccine