Proteína capsidial do Rupestris stem pitting-associated vírus: seqüenciamento do gene, expressão em Escherichia coli, purificação e produção de anti-soro policlonal

摘要: PEREIRA, A.C.B. Capsid protein of Rupestris stem pitting-associated virus: sequencing the gene, expression in E. coli, purification and production polyclonal antiserun. Sao Jose do Rio Preto, 2008. 66p. Dissertacao (Mestrado em Microbiologia) – Instituto de Biociencias, Letras e Ciencias Exatas, Universidade Estadual Paulista “Julio Mesquita Filho”. pitting (RSP), a component rugose wood (RW) complex, is one most graft-transmissible grapevine virus diseases with great economic importance for viticulture . (RSPaV), genus Foveavirus within family Flexiviridae, has been associated this disease. This work reports coat (CP) gene brazilian an isolate RSPaV (RSPaV-SP), its Escherichia recombinant antiserum rabbit. CP was found to be 780nt long, 256 deduced amino acid sequence encoding predicted 28 kDa. In filogenetic analysis, RSPaV-SP other variants virus, four groups were showed highest identity variant RSPaV-BS. The purified by affinity chromatography molecular weight 32kDa (4 kDa from small vector plus RSPaV-SP). proved specific detection Western Blot, did not react heterologous proteins starting at dilution 1:4000. At dilution, effective leaf extracts infected plants no reaction observed healthy grapevines. Considering that low concentrations during seasons year, results obtained so far highly satisfactory detection. Serological methods have advantages over biological indexing method, since they are cheaper can used large-scale tests such as ELISA. Experiments using ELISA technique successful. Purification native would alternative more efective detect these technique.