Produção das proteínas recombinantes AM254, VirB9 e VirB10, de Anaplasma marginale e avaliação preliminar de sua antigenicidade. / Production of recombinant proteins AM254, VirB9 and VirB10 of Anaplasma marginale and preliminary evaluation its antigenicity.

AUTOR(ES)

Cátia Marques da Costa

DATA DE PUBLICAÇÃO

2007

RESUMO

Anaplasma marginale (Ricktsialles: Anaplasmataceae) is a ricketsial hemoparasite responsible for causing great economic losses in cattle from tropical and subtropical regions. The objectives of this work were to produce the recombinant membrane proteins AM254, VirB9 and VirB10 and to evaluate its possible antigenicity. The genes am254, virB9 and virB10 were submitted to various experiments and were , linked to pET47 (b) plasmid. After the certification of the recombinant plasmid (pET-47-am254, pET-47-virB9 and pET-47- virB10), it was transformed into E. coli Rosetta cells for expression. The recombinant proteins produced, were analyzed by Western blot assay where the reaction of the anti-histidin monoclonal antibody against rAM254 (47kDa), rVirB9 (31kDa) and rVirB10 (60kDa) was observed. After the confirmation of the production of recombinant proteins the indirect Enzyme-Linked Immnunosorbent Assay (ELISA) was standardized with sera previously confirmed by PCR and ELISA (rMSP1 and rMSP5). The averages of the optic densities (OD) of the positive sera were of 1.339; 1.288 and 1.240 and of the negative sera of 0.470, 0.324 and 0.414 for AM254, VirB9 and VirB10 respectively with significant difference to the level of 5% between positives and negatives for each recombinant protein. The study demonstrated that the proteins rAM 254, rVirB9 and rVirB10 of A. marginale are recognized by sera of bovines immunized by different Brazilian isolates of the ricketsia (homologous and heterologous), showing the antigenicity potential of these proteins.

ASSUNTO(S)

proteínas recombinantes de membrana medicina veterinaria recombinant membrane protein diagnóstico. anaplasmataceae diagnostic.

Documentos Relacionados

• Interactions between VirB9 and VirB10 membrane proteins involved in movement of DNA from Agrobacterium tumefaciens into plant cells.
• IgG and IgG2 antibodies from cattle naturally infected with Anaplasma marginale recognize the recombinant vaccine candidate antigens VirB9, VirB10, and elongation factor-Tu
• The Agrobacterium T-DNA Transport Pore Proteins VirB8, VirB9, and VirB10 Interact with One Another
• Complementation analysis of Agrobacterium tumefaciens Ti plasmid virB genes by use of a vir promoter expression vector: virB9, virB10, and virB11 are essential virulence genes.
• Agrobacterium tumefaciens VirB7 and VirB9 form a disulfide-linked protein complex.