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Doctoral Thesis
DOI
https://doi.org/10.11606/T.11.1990.tde-20210104-182731
Document
Author
Full name
Maria das Gracas Ongarelli
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
Piracicaba, 1990
Supervisor
Title in Portuguese
Aplicação da técnica de eletroforese na caracterização de fragmentos de restrição e proteínas dos baculovírus de Anticarsia gemmatalis Hueb., 1818 (Lepidoptera: Noctuidae) e Diatraea saccharalis Fabr., 1794 (Lepidoptera: Pyralidae)
Keywords in Portuguese
ÁCIDOS NUCLEICOS
BACULOVIRUS
BROCA DA CANA-DE-AÇÚCAR
ELETROFORESE
LAGARTA DA SOJA
PROTEINAS
Abstract in Portuguese
No presente trabalho foram desenvolvidos estudos sobre a natureza nucleoproteica do vírus da poliedrose nuclear de Anticarsia gemmatalis Hueb, 1818 (AgNPV) e o vírus da granulose de Diatraea saccharalis Fahr., 1794 (DsGV) com a utilização de técnicas que pudessem discriminar diferenças entre esses baculovírus. Fez-se um estudo prévio do processo mais conveniente de extração dos ácidos nucleicos com base na relação de densidade óptica a 260/280 nm. Os tratamentos dos ácidos nucleicos do AgNPV com Eco RI e DsGV com Bsp RI produziram fragmentos de restrição caracterizados no perfil eletroforético em agarose com a determinação dos pesos moleculares. Eco RI produziu 6 fragmentos de restrição no DNA de AgNPV e Bsp RI produziu 4 fragmentos de restrição no DNA de DsGV. Foram obtidos resultados preliminares da possível caracterização de DNAs dos baculovírus através da técnica de "Finger-print" dos ácidos nucleicos com enzimas de restrição. As proteínas do AgNPV e do DsGV após sofrerem extração foram corridas em gel SDS - poliacrilamida produzindo perfis eletroforéticos com 22 bandas para o AgNPV e 18 bandas para o DsGV para os quais foram determinados os respectivos pesos moleculares. Foi possível identificar nos perfis de proteínas a poliedrina, a granulina e a proteína 39K que são sintetizadas em grande quantidade pelas células infectadas. A proteína p10 não foi encontrada nos perfis analisados. Foram confrontadas diferenças entre os pesos moleculares das proteínas do DsGV, AgNPV e AcNPV (Autographa californica). Levando-se em conta que as divergências entre os vírus da granulose e da poliedrose nuclear são mais acentuadas que entre as granuloses e as poliedroses entre si é possível que as proteínas heterólogas do DsGV (44,4%) e as do AcNPV (40,9%) com relação as proteínas do AgNPV sejam significativas.
Title in English
Electrophoresis technique application for the characterization of restriction fragments and proteins of the baculovirus of Anticarsia gemmatalis Hueb., 1818 (Lepidoptera: Noctuidae) and Diatraea saccharalis Fabr., 1794 (Lepidoptera: Pyralidae)
Keywords in English

Abstract in English
Studies have been carried out on the nucleoproteic nature of the nuclear polyhedrosis virus of the Anticarsia gemmatalis (Hueb, 1818) (AgNPV) and the granulosis virus of Diatraea saccaharalis (Fabr., 1794) (DsGV) using techniques that would help discriminate differences between the two baculoviruses. The most common process of nucleic acid extraction has been previously studied based on optical density relation at 260 and 280 nm. Treatment of the nucleic acids of AgNPV with Eco RI and of DsGV with Bsp RI produced restriction fragments characterized in the electrophoretic profile in agarose with the determination of the molecular weights. Eco RI produced 6 (six) restriction fragments in AgNPV-DNA and Bsp RI produced 4 (four) restriction fragments in DsGV-DNA. Preliminary results of the possible characterization of the DNA's of the baculoviruses were observed by using the "Finger-print" technique of nucleic acid with restriction enzymes. The proteins of AgNPV and DsGV after extraction were runned in polyacrilamide - SDS - gel producing profiles with 22 bands for the AgNPV and 18 bands for the DsGV, of which the respectives molecular weights were determined. It was possible to identify on the protein profiles the polyhedrin, the granulin and the 39K protein, which are synthesized in large amounts by the infected cells. The p10 protein was not found in the profiles analyzed. The differences among the molecular weights of the proteins of DsGV, AgNPV and AcNPV (Autographa californica) were compared. Taking into account that the divergences between the granulosis virus and polyhedrosis nuclear virus are more evident than among the polyhedrosis and granulosis viruses, it is possible that the heterologous proteins of DsGV (44.4%) and AcNPV (40.9%) in relation to the proteins of AgNPV might be significant.
 
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Publishing Date
2021-01-07
 
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