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Master's Dissertation
DOI
https://doi.org/10.11606/D.11.1986.tde-20220207-234844
Document
Author
Full name
Luís Otávio Saggion Beriam
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Piracicaba, 1985
Supervisor
Title in Portuguese
Purificação simplificada do vírus do enrolamento da folha da batata com triton X-100
Keywords in Portuguese
BATATA
ENROLAMENTO DA FOLHA
POLEROVIRUS
PURIFICAÇÃO
Abstract in Portuguese
Várias metodologias foram empregadas para purificação do vírus do enrolamento da folha da batata (Potato Leaf-roll Virus PLRV). Purificações com o emprego de clorofórmio/butanol, polietilenoglicol e Triton X-100 foram efetuadas. A técnica de Triton X-100 se mostrou mais eficiente para purificação do VEFB. Tal técnica consistiu no tratamento de plantas infectadas com uma solução de Triton X-100 a 2.5% em tampão fosfato (KH2PO4) 0,5M pH 7, seguido de um ciclo de centrifugações diferenciais e gradiente de densidade em sacarose. Os precipitados resultantes foram observados ao microscópio eletrônico, revelando a presença de partículas do VEFB. O uso de Triton X-100 abre uma nova perspectiva para a purificação de fitovírus. É importante ressaltar que se trata de purificação do VEFB com o uso exclusivo de Triton X-100, sem o emprego de solventes orgânicos ou quaisquer outras substâncias comumente empregadas no processo de purificação de vírus. Antissoro para o VEFB foi obtido através da técnica de imunização via linfonódulo, apresentando um título de 1:2.
Title in English
Not available
Abstract in English
Some purification methods were employed to purify the Potato Leafroll Virus ( PLRV). Purifications utilizing chloroform/butanol, polyethileneglicol and Triton X-100 were made. The Triton X-100 technique was efficient to purify the PLRV. The infected plants were treated with Triton X-100 at 2,5% in, buffer KH2PO4 pH 7 and submitted to a cicle of differential centrifugations and density sucrose gradient. The pellets were observed in electron microscopy and virus particles were seen. The use of Triton X-100 opens up a new perspective for the purification of fitovirus. It's important to note that this deals with the purification of PLRV with the exclusive use of Triton X-100, without the use of organic solvents or other substances normally employed in the virus purification. Antiserum to PLRV were prepared by the lymph node technique immunization, showing a titre of 1:2.
 
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Publishing Date
2022-02-07
 
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