RESUMEN
A diagnostic kit for detection of avian influenza virus by real-time polymerase chain reaction was developed. This kit allows to identify the influenza viruses type A and highly pathogenic strain of avian influenza virus H5N1. The diagnostic kit is universal and adopted for ABI PRISM SDS (Applied Biosystems), RotorGene (Corbett Research) and iQCycler (BioRad) PCR machines.
Asunto(s)
Subtipo H5N1 del Virus de la Influenza A , Gripe Aviar/virología , Reacción en Cadena de la Polimerasa/métodos , Secuencia de Aminoácidos , Animales , Aves , Subtipo H5N1 del Virus de la Influenza A/genética , Subtipo H5N1 del Virus de la Influenza A/aislamiento & purificación , Subtipo H5N1 del Virus de la Influenza A/patogenicidad , Datos de Secuencia Molecular , ARN Viral/genética , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , VirulenciaRESUMEN
Hybridization analysis of the double-strand RNA-elements isolated from the sugar beet of the Kyiv, Vinnytsya and Poltava Regions was carried out. A positive signal has been obtained with the dsRNA fragment, 1.9 kbp long, which codes the viral RNA- dependent RNA-polymerase having high level of homology with the analogous enzyme of mycoviruses from Partitiviridae family. Molecular identity of these dsRNA-elements in the sugar beet samples from the Vinnytsya and Poltava regions has been shown The RT-PCR test system has been developed for detecting the nondescribed mycovirus which is proposed to be called Helicobasidium purpureum partitivirus.
Asunto(s)
Beta vulgaris/virología , Virus de Plantas/genética , Virus ARN/genética , Virus ARN/aislamiento & purificación , ARN Bicatenario/genética , Secuencia de Aminoácidos , Secuencia de Bases , Secuencia Conservada , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Virus de Plantas/química , Virus ARN/clasificación , ARN Bicatenario/química , ARN Viral/genética , ARN Polimerasa Dependiente del ARN/química , ARN Polimerasa Dependiente del ARN/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Homología de Secuencia de Aminoácido , Homología de Secuencia de Ácido NucleicoRESUMEN
Localization of virus RNA in stroma of Capsicum anuum L. chloroplasts was determined by the PCR method. Accumulation of virus protein in the membranes and stroma of infected pepper chloroplasts has been studied. It is concluded that the virus protein synthesis takes place in the pepper chloroplasts.
Asunto(s)
Capsicum/virología , Cloroplastos/virología , Reacción en Cadena de la Polimerasa/métodos , ARN Viral/ultraestructura , Capsicum/anatomía & histología , Cloroplastos/patología , Cloroplastos/ultraestructura , Virus del Mosaico del Tabaco/ultraestructuraRESUMEN
Chlorotic areas of Capsicuum anuum L. leaves infected with tobacco mosaic virus (TMV) have been investigated. Pathological changes of chloroplasts were found out. Swelling, more osmophilic plastoglobuli, loosened thylakoid structure were observed. It was shown that chloroplasts did not take part in the process of virus replication. Some structural changes of mitochondria were discovered too. They were most tolerant to the virus infection in comparison with other organoids.
Asunto(s)
Capsicum/virología , Enfermedades de las Plantas/virología , Hojas de la Planta/virología , Virus del Mosaico del Tabaco/patogenicidad , Capsicum/anatomía & histología , Cloroplastos/patología , Cloroplastos/ultraestructura , Cloroplastos/virología , Edema/patología , Edema/virología , Mitocondrias/patología , Mitocondrias/ultraestructura , Mitocondrias/virología , Hojas de la Planta/ultraestructura , Tilacoides/patología , Tilacoides/ultraestructura , Tilacoides/virología , Replicación ViralRESUMEN
We used yeast three-hybrid system, for studying interaction of alfalfa mosaic virus coat protein AMVCP (AMVCP) with RNA4, which codes this protein. We have shown that AMVCP with high affinity is bound to plus-chain of RNA4 in vivo. The mutational analysis has shown, that the N-terminal part of AMVCP (aa 1 to 85) contains RNA-binding domain. C-terminal part of this protein (aa 86 to 221) does not participate in direct interaction with RNA4. However activity of the reporter-gene LacZ, which codes beta-galactosidase, in case of interaction only N-terminal part of AMVCP is five times lower, in comparison with full-length hybrid protein, that confirms that the tertiary structure of full-length AMVCP is more favourable for interaction with RNA4.