Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 3 de 3
Filtrar
Mais filtros

Base de dados
Ano de publicação
Tipo de documento
Intervalo de ano de publicação
1.
Vet Parasitol ; 190(3-4): 401-10, 2012 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-22819586

RESUMO

The Babesia bigemina heat shock protein-70 gene (BbigHSP-70) was cloned from cDNA by polymerase chain reaction (PCR) and sequenced. The length of the gene is 1947 bp and the predicted polypeptide is 649 amino acids long with a calculated molecular weight of 70.85 kDa. BbigHSP-70 has a signal peptide of 15 amino acids. Phylogenetic analysis of the amino acid sequence of BbigHSP-70 showed that B. bigemina was most closely related to B. caballi and B. bovis and lies within a phylogenetic cluster with Theileria. rBbigHSP-70 was expressed in E. coli as a soluble GST-fusion protein with a molecular mass of 96.8-kDa. The serum raised in mice against rBbigHSP-70 detected the native protein in B. bigemina, B. bovis, B. caballi, B. gibsoni, and B. microti lysates and also reacted with B. bigemina, B. bovis, and B. caballi merozoites in the IFAT. Mice vaccinated with rBbigHSP-70 showed lower parasitemia against the challenge infection with B. microti than GST-vaccinated and non-vaccinated controls. These results added a new member of Babesia heat shock proteins70 that is well conserved among intraerythrocytic protozoa and demonstrated its protective effects in an experimental model of rodent babesiosis.


Assuntos
Babesia/metabolismo , Regulação da Expressão Gênica/fisiologia , Proteínas de Choque Térmico HSP70/metabolismo , Sequência de Aminoácidos , Animais , Babesia/genética , DNA de Protozoário/genética , DNA de Protozoário/metabolismo , Feminino , Proteínas de Choque Térmico HSP70/genética , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Filogenia , RNA de Protozoário/genética , RNA de Protozoário/metabolismo
2.
Mol Biochem Parasitol ; 178(1-2): 40-5, 2011.
Artigo em Inglês | MEDLINE | ID: mdl-21406202

RESUMO

A cDNA encoding a new Babesia bovis spherical body protein 4 (BbSBP-4) was reported to have no significant homology to other apicomplexan proteins or previously reported B. bovis spherical body proteins. In the present study, we further examined the molecular characteristics of BbSBP-4 including the expression and cellular localization of the BbSBP-4. An anti-rBbSBP-4 mouse serum specifically reacted to a 41-kDa native protein B. bovis in Western blot analysis. The immunoelectron microscopic examination confirmed the localization of BbSBP-4 in spherical bodies, but not in the nucleus, rhoptries, and micronemes. Interestingly, the protein was found to be localized not only in the spherical body of B. bovis but also in the cytoplasm of infected erythrocytes (iRBC) at the later stage of parasite development. The confocal laser microscopic examination and Western blot analysis demonstrated the increased accumulation of BbSBP-4 in the cytoplasm of iRBC and in the supernatant of cultivated B. bovis during the late developmental stage of the parasite. These results suggest that BbSBP-4 was secreted from spherical body into cytoplasm of iRBC during the late developmental stage of the parasite before the rupture of infected RBC. Taken together, BbSBP-4 might play an important role as a secreted protein in the intracellular development and/or survival of B. bovis.


Assuntos
Babesia bovis/metabolismo , Eritrócitos/parasitologia , Proteínas de Protozoários/metabolismo , Animais , Western Blotting , Citoplasma/química , Citoplasma/parasitologia , DNA de Protozoário/química , DNA de Protozoário/genética , Camundongos , Microscopia Confocal , Microscopia Imunoeletrônica , Dados de Sequência Molecular , Organelas/química , Proteínas de Protozoários/genética , Análise de Sequência de DNA
3.
Clin Vaccine Immunol ; 18(2): 337-42, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21123520

RESUMO

Five Babesia bovis recombinant proteins, including merozoite surface antigen 2c (BbMSA-2c), C-terminal rhoptry-associated protein 1 (BbRAP-1/CT), truncated thrombospondin-related anonymous protein (BbTRAP-T), spherical body protein 1 (BbSBP-1), and spherical body protein 4 (BbSBP-4), were evaluated as diagnostic antigens to detect the infection in cattle. The recombinant proteins were highly antigenic when tested with experimentally B. bovis-infected bovine serum in Western blot analysis. Furthermore, five antisera that had been raised against each of the recombinant proteins reacted specifically with the corresponding authentic protein, as determined in Western blot analysis. Next, enzyme-linked immunosorbent assays (ELISAs) using these recombinant proteins were evaluated for diagnostic use, and the sensitivity and specificity of each protein were demonstrated with a series of serum samples from experimentally B. bovis-infected cattle. Furthermore, a total of 669 field serum samples collected from cattle in regions of B. bovis endemicity in seven countries were tested with the ELISAs, and the results were compared to those of an indirect fluorescent antibody test (IFAT), as a reference. Among five recombinant antigens, recombinant BbSBP-4 (rBbSBP-4) had the highest concordance rate (85.3%) and kappa value (0.705), indicating its reliability in the detection of specific antibodies to B. bovis in cattle, even in different geographical regions. Overall, we have successfully developed an ELISA based on rBbSBP-4 as a new serological antigen for a practical and sensitive test which will be applicable for epidemiologic survey and control programs in the future.


Assuntos
Anticorpos Antiprotozoários/sangue , Antígenos de Protozoários , Babesia bovis/isolamento & purificação , Babesiose/veterinária , Doenças dos Bovinos/diagnóstico , Parasitologia/métodos , Proteínas de Protozoários , Animais , Antígenos de Protozoários/genética , Babesia bovis/imunologia , Babesiose/diagnóstico , Babesiose/parasitologia , Western Blotting , Bovinos , Doenças dos Bovinos/parasitologia , DNA de Protozoário/química , DNA de Protozoário/genética , Ensaio de Imunoadsorção Enzimática/métodos , Dados de Sequência Molecular , Proteínas de Protozoários/genética , Proteínas Recombinantes/genética , Sensibilidade e Especificidade , Análise de Sequência de DNA
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA