Cloning and expression of a biotinylated multiple-epitope HCV fusion antigen gene / 中国实验血液学杂志
Journal of Experimental Hematology
;
(6): 359-362, 2004.
Artigo
em Chinês
| WPRIM
| ID: wpr-352065
ABSTRACT
The aim was to develop a single multiple-epitope fusion antigen which incorporates all of the major immunodominant epitopes from the six functional regions of the HCV genome. A nucleic acid sequence consisting of viral core, E1, E2, NS3, NS4, and NS5 regions was constructed and inserted into the Promega Pinpoint Xa-1 T vector for inducing expression. The protein was expressed in JM109 (DE3) as a fusion protein with a 13 kD biotinlated tag to be used for detection and affinity purification. Immunogenicity and biotinylated tag of the fusion protein were detected by Western blot analysis with positive anti-HCV serum and streptavidin alkaline phosphatase. After purified by Promega SoftLink Soft Release Avidin Resin, the protein was pre-coated on microwell and detected with anti-core, anti-NS3, anti-NS4 and anti-NS5 positive sera by EIA, respectively. The results indicated that the recombinant soluble protein was expressed and labelled with biotin successfully, it reacted with anti-HCV positive serum, and exposed all of the major immunogenic epitopes chosen. In conclusion, this recombinant antigen may be used to design an double antigen sandwich anti-HCV immunoassay.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Sangue
/
Proteínas Recombinantes de Fusão
/
Ensaio de Imunoadsorção Enzimática
/
Epitopos Imunodominantes
/
Clonagem Molecular
/
Hepacivirus
/
Anticorpos Anti-Hepatite C
/
Biotinilação
/
Alergia e Imunologia
/
Genética
Limite:
Humanos
Idioma:
Chinês
Revista:
Journal of Experimental Hematology
Ano de publicação:
2004
Tipo de documento:
Artigo
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