RESUMO
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.
Assuntos
Virus da Influenza A Subtipo H5N1 , Influenza Aviária/virologia , Reação em Cadeia da Polimerase/métodos , Sequência de Aminoácidos , Animais , Aves , Virus da Influenza A Subtipo H5N1/genética , Virus da Influenza A Subtipo H5N1/isolamento & purificação , Virus da Influenza A Subtipo H5N1/patogenicidade , Dados de Sequência Molecular , RNA Viral/genética , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , VirulênciaRESUMO
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.
Assuntos
Beta vulgaris/virologia , Vírus de Plantas/genética , Vírus de RNA/genética , Vírus de RNA/isolamento & purificação , RNA de Cadeia Dupla/genética , Sequência de Aminoácidos , Sequência de Bases , Sequência Conservada , Dados de Sequência Molecular , Hibridização de Ácido Nucleico , Vírus de Plantas/química , Vírus de RNA/classificação , RNA de Cadeia Dupla/química , RNA Viral/genética , RNA Polimerase Dependente de RNA/química , RNA Polimerase Dependente de RNA/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido NucleicoRESUMO
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.
Assuntos
Proteínas do Capsídeo/metabolismo , RNA Viral/metabolismo , Proteínas de Ligação a RNA/metabolismo , Western Blotting , Proteínas do Capsídeo/genética , Clonagem Molecular , Genes Reporter/genética , Óperon Lac/genética , Ligação Proteica , Estrutura Terciária de Proteína , RNA Viral/genética , Proteínas de Ligação a RNA/genética , Saccharomyces cerevisiae/genética , Transcrição Gênica , Técnicas do Sistema de Duplo-Híbrido , beta-Galactosidase/genética , beta-Galactosidase/metabolismoRESUMO
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.
Assuntos
Capsicum/virologia , Cloroplastos/virologia , Reação em Cadeia da Polimerase/métodos , RNA Viral/ultraestrutura , Capsicum/anatomia & histologia , Cloroplastos/patologia , Cloroplastos/ultraestrutura , Vírus do Mosaico do Tabaco/ultraestruturaRESUMO
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.
