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Doctoral Thesis
DOI
https://doi.org/10.11606/T.42.2010.tde-25032010-160746
Document
Author
Full name
Fernanda Perez Yeda
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2010
Supervisor
Committee
Harsi, Charlotte Marianna (President)
Mehnert, Dolores Ursula
Mendonça, Ronaldo Zucatelli
Pereira, Luiz Eloy
Ventura, Armando Morais
Title in Portuguese
Montagem de um pseudo-hantavírus quimera, contendo a nucleoproteína do vírus Araraquara e as glicoproteínas do vírus Andes, em sistema baculovírus
Keywords in Portuguese
Expressão de proteínas
Glicoproteínas
Nucleoproteína
Proteínas
Sistema baculovírus
Vírus
Abstract in Portuguese
Os hantavírus, membros da família Bunyaviridae, são os agentes infecciosos responsáveis pela Febre Hemorrágica com Síndrome Renal e pela Síndrome Cardiopulmonar por Hantavírus. São vírus com genoma constituído por três segmentos de RNA fita simples, de polaridade negativa, designados como S, M e L, que codificam, respectivamente, a nucleoproteína, as glicoproteínas G1 e G2 e a RNA polimerase dependente de RNA. Com o objetivo de estudar a montagem de pseudopartículas quiméricas de hantavírus, a proteína N do vírus Araraquara e as glicoproteínas G1 e G2 do vírus Andes foram expressas em sistema baculovírus. A microscopia confocal mostrou a colocalização das proteínas G1 e G2 com a proteína N. Pelos ensaios de imunoprecipitação e de centrifugação em gradiente de sacarose, foi observada a interação entre as proteínas N, G1 e G2. Nas análises por microscopia eletrônica de transmissão foi observada a montagem do pseudo-hantavírus quimera, com morfologia semelhante ao do vírion. O pseudo-hantavírus quimera obtido neste estudo poderá, no futuro, ser utilizado em estudos imunológicos, estruturais e morfológicos.
Title in English
Assembly of a chimeric hantavirus-like particle, containing the Araraquara nucleoprotein and the Andes glycoproteins, expressed in baculovirus system
Keywords in English
Baculovirus system
Glycoprotein
Nucleoprotein
Protein
Protein expression
Virus
Abstract in English
Hantaviruses, members of the Bunyaviridae family, are the infectious agents responsible for Hemorrhagic Fever with Renal Syndrome and the Hantavirus Cardiopulmonary Syndrome. The viral genome is composed by three segments of single-stranded negative-sense RNA, designated as S, M and L, which encode, respectively, the nucleoprotein, the G1 and G2 glycoproteins, and the RNA-dependent RNA polymerase. In order to study the assembly of a chimeric hantavirus-like particle, the Araraquara nucleoprotein and the Andes glycoproteins were expressed in a baculovirus system. Confocal microscopy showed the colocalization of G1 and G2 proteins with the N protein. Immunoprecipitation assay and sucrose density gradient showed the interaction among N, G1 and G2 proteins. The transmission electron microscopy showed the hantavirus-like particle with the same morphology of the virion. The chimeric hantavirus-like particle produced in this study could be used, in the future, in immunological, structural and morphological studies.
 
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Publishing Date
2010-03-29
 
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