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1.
Journal of Veterinary Science ; : 149-157, 2001.
Artículo en Inglés | WPRIM | ID: wpr-109440

RESUMEN

The targeted RNA recombination was attempted to substitute the membrane (M) protein gene and part of the nucleocapsid (N) protein gene of mouse hepatitis virus with the corresponding sequences from bovine coronavirus. Using a defective interfering (DI) RNA-like cDNA construct derived from pMH54, 690 nucleotides representing the entire M gene and the 5' most 915 nucleotides of the N gene of the mouse hepatitis virus Albany 4 mutant were attempted to be replaced. Upon infection of cells with Albany 4 followed by transfection with synthetic RNA transcribed from the DI-like cDNA construct, recombinant mouse hepatitis viruses as the large plaque forming phenotype were isolated by plaque assays at the non-permissive temperature of 391 degrees C. By RT-PCR and sequencing, those large plaque phenotypes were confirmed to have contained the thermostable phenotype marker derived from the transfected RNA, demonstrating that recombination occurred between the Albany 4 genomic RNA and the in vitro RNA transcripts. Further analysis of the recombinant viruses indicated that there combination had taken place within the region of 222 nucleotides between positions 916 and 1,137 of the N gene. This is the region immediately downstream of the replacement sequence and the start of the temperature resistant phenotype marker. The results suggest that the M and part of the N genes of bovine coronavirus may not be able to complement the function of those of mouse hepatitis virus. This study redirects our current approach of utilizing the MHV targeted RNA recombination as a means to study bovine coronavirus genetics towards the construction of an infectious cDNA clone.


Asunto(s)
Animales , Bovinos , Ratones , Secuencia de Aminoácidos , Secuencia de Bases , Células Cultivadas , Coronavirus Bovino/genética , ADN Complementario/genética , Marcación de Gen/veterinaria , Vectores Genéticos , Datos de Secuencia Molecular , Virus de la Hepatitis Murina/genética , Proteínas de la Nucleocápside/genética , Fenotipo , Ensayo de Placa Viral/veterinaria , ARN Viral/química , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Homología de Secuencia de Aminoácido , Transfección/veterinaria , Proteínas de la Matriz Viral/genética
2.
Southeast Asian J Trop Med Public Health ; 1995 Mar; 26(1): 109-13
Artículo en Inglés | IMSEAR | ID: sea-34149

RESUMEN

A study was conducted to measure the prevalence of hemagglutination-inhibition (HI) and neutralizing antibodies against two arboviruses (Chikungunya and Japanese encephalitis virus) in horses of Java, Indonesia. Blood specimens were collected from a sample of 112 horses at two stables: Pulo Mas, a racing track-horse complex, located in a residential area in North Jakarta, and Pamulang, a riding school, located in a rural environment of West Jaya. Sera were tested by the HI assay and plaque reduction neutralization test. JEV antibodies were detected by HI in 58 (52%) of the horses, while only 11 (10%) had Chikungunya antibodies by HI. The proportion of Pamulang horses infected with JEV (66%) was significantly higher than found among Pulo Mas horses (40%) screened (p < 0.01). Of the 58 horses with JEV antibodies by HI, 52 (90%) were found to have specific neutralization antibodies to JEV. HI and neutralization tests on horse sera indicated that the risk to alpha virus infections was minimal in horses surveyed from Java. However, there was a high risk of JEV infection among the same population.


Asunto(s)
Infecciones por Alphavirus/prevención & control , Animales , Anticuerpos Antivirales/análisis , Virus Chikungunya/inmunología , Reservorios de Enfermedades/veterinaria , Virus de la Encefalitis Japonesa (Especie)/inmunología , Encefalitis Japonesa/prevención & control , Pruebas de Inhibición de Hemaglutinación/veterinaria , Enfermedades de los Caballos/epidemiología , Indonesia , Pruebas de Neutralización/veterinaria , Ensayo de Placa Viral/veterinaria , Prevalencia
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