Cloning, expression and characterization of an antigenic domain of the a major surface protein recombinant NcSRS2 (Nc1-p43) of Neospora caninum. / Clonagem, expressão e caracterização do domínio antigênico da proteína majoritária de superfície NcRS2 (Nc1-p43) recombinante de Neospera caninum

AUTOR(ES)

Manoel Sebastião da Costa Lima Junior

DATA DE PUBLICAÇÃO

2006

RESUMO

Neospora caninum is an obligate intracellular protozoan that can infect domestic and wild canids, as well as ruminants and equines. It was described in 1988 as causing neuromuscular alterations and death in dogs. Recently, N. caninum has received considerable attention for its large impact on the dairy industry, given the economic losses related to breeding failures and to a decrease in productivity. ELISA diagnosis of neosporosis has not been widely used in Brazil, mostly because of the cost of assay, and thus the distribution of the disease in the country is still not well known. In order to evaluate its ability to react with sera from infected animals from the state of Mato Grosso do Sul, an antigenic determinant domain of a major surface protein (Nc-p43) was produced. The antigenic domain, located in the distal 2/3 region of the C-terminus, was amplified by polymerase chain reaction. The DNA fragments were cloned into pet100/D-TOPO vectors. The recombinant plasmids were transformed into Escherichia coli of the BL21 Star (DE3) strain and induced to express the fused fragment of Nc-p43 as a 29-kDa protein that, when assayed with bovine Neospora-positive serum from a regional sample, was sensitive for identification by immunoblotting. This fragment of Nc-p43 may have use in additional studies targeted at diagnosing N. caninum infection and at evaluating the immunoprotection conferred by the protein fragment to animal hosts.

ASSUNTO(S)

biologia molecular nc-p43 antigen recombinant protein proteína recombinante neospora caninum antígenos

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