Expression of tick-borne encephalitis virus prM-E protein in insect cells and studies on its antigenicity / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
;
(6): 335-339, 2005.
Artigo
em Chinês
| WPRIM
| ID: wpr-333011
ABSTRACT
<p><b>BACKGROUND</b>To express the prM-E protein in Sf9 cells, and lay a basis for further study on the function of the viral proteins and development of specific diagnostic reagents.</p><p><b>METHODS</b>The recombinant prM-E protein of tick-borne encephalitis virus was expressed in insect cell Sf9 by RT-PCR amplification of prM-E gene, construction of donor plasmid of Bac-to-Bac baculovirus expression system, homologous recombination of donor plasmid with bacmid DNA at the site of Tn7 and transfection of insect cell Sf9.</p><p><b>RESULTS</b>Recombinant subviral particles, about 30 nm in diameter, consisting of prM-E were observed by electron microscope in the supernatant of infected cells, which indicated that infected cells released virus-like particles (VLPs) into the culture medium. The results of Western-blot and the indirect immunofluorescence assay (IFA) showed that the recombinant proteins retained antigenic and conformational structures similar to those of native virus proteins. Using the recombinant prM-E as antigens to detect samples of patient sera by ELISA and IFA, all of 16 sera from patients with tick-borne encephalitis were positive and all of 6 sera from other patients were negative.</p><p><b>CONCLUSION</b>The prM-E protein expressed in insect cells retains good antigenicity.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Regulação Viral da Expressão Gênica
/
Linhagem Celular
/
Proteínas do Envelope Viral
/
Western Blotting
/
Spodoptera
/
Técnica Indireta de Fluorescência para Anticorpo
/
Reação em Cadeia da Polimerase Via Transcriptase Reversa
/
Vírus da Encefalite Transmitidos por Carrapatos
/
Alergia e Imunologia
/
Genética
Limite:
Animais
Idioma:
Chinês
Revista:
Chinese Journal of Experimental and Clinical Virology
Ano de publicação:
2005
Tipo de documento:
Artigo
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