Expressão, purificação e caracterização de proteínas dos fitovírus SBMV e RSPaV

• Autor(a) principal: Mariutti, Ricardo Barros [UNESP]
• Data de Publicação: 2014
• Tipo de documento: Tese
• Idioma: por
• Título da fonte: Repositório Institucional da UNESP
• Texto Completo: http://hdl.handle.net/11449/127647; http://www.athena.biblioteca.unesp.br/exlibris/bd/cathedra/14-09-2015/000846547.pdf
• topic: Fitopatologia; Virus de plantas; Videira; Proteínas virais - Purificação

The grapevine culture is the most economically important fruit plant in the world. The grapevine is the basic source of high valued wines and other alcoholic beverages. The Rupestris stem pitting associated virus-RSPaV was identified as the causative agent of the rupestris stem pitting (RSP) disease, a component ofthe the rugose wood (RW) complex, one of the major disease complexes affecting grapevines. This disease is transmitted by grafting and is one of the most widespread viruses of grapevine in the world. In brazil this disease was reported for the first time in late 1960s. As it is difficult to seperate the viral particles from the infected plant, the expression of large-scale of coat protein of RSPaV in bacteria is a promising strategy for understanding their properties. The bean crop productivity has also greatly affected by the occurrence of diseases that reduce the production and consequently reduce their supply, causing an increasein the prices. This virus has a narrow host range, confined almost exclusively to species of leguminous species, some of the species has a very high economical value such as the common bean and soybean. The movement protein (MP), serine protease and VPg, are produced in low concentrations, making impossible the large-scale purification from infected plants. In Brazil, more than ten virus were reported in beans, among them, Southern bean mosaic virus (SBMV) from the genus Sobemovirus. The large scale expression and purification of proteins VPg, serine protease and movement protein of SBMV and capisidial protein of RSPaV were performed using pMAL pET28a vectors. Analysis of the proteins by dynamic light scattering showed that was possible to obtain monomodal protein solutions. By circular dichroism was observed that the proteins were structured and the investigation of protein-ligand interactions was performed using nuclear magnetic resonance. These results contributed to increase the knowledge...