The expression and bioactivity of S gene in Vero-E6 cells using baculovirus vector / 中华实验和临床病毒学杂志
Chinese Journal of Experimental and Clinical Virology
; (6): 280-282, 2011.
Article
em Zh
| WPRIM
| ID: wpr-246265
Biblioteca responsável:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>The S gene of a Hanta Virus (HV) Z10 strain was cloned into a baculovirus shuttle bacmid pDual-CMV contained a CMV promoter to generated a recombinant baculovirus BAC-pDual-CMV-HVS, then the recombinant baculovirus was transfected into Vero-E6 cell. The cells with recombinant baculovirus were applied to the detection of HV antiserum.</p><p><b>METHODS</b>To generate the recombinant baculovirus BAC-pDual-CMV-HVS, the sequence of CMV promoter was obtained from the plasmid pEGFP-N1 by PCR, and subsequently cloned to the baculovirus shuttle bacmid pFastBacDUAL resulting the recombinant plasmid pDual-CMV. Then the sequence of HV-S gene was inserted to the plasmid pDual-CMV, to generate the plasmid pDual-CMV-HVS. Plasmid pDual-CMV-HVS was transformed into the DH10BAC competent cells to get the recombinant baculovirus BAC-pDual-CMV-HVS. The antigen substrate slides were made by transfecting the recombinant virus into Vero-E6 cells.</p><p><b>RESULTS</b>The plasmid pDual-CMV-HVS was verified by sequencing. The recombinant virus BAC-pDual-CMV-HVS was generated according to the protocol of the baculovirus and transfected into Vero-E6 cells. The expression of the HV-S gene was verified by positive HV antiserum.</p><p><b>CONCLUSION</b>[corrected] The recombinant virus were successfully generated and applied to prepare the antigen substrate slides. The antigen substrate slides was conveniently prepared without special equipments, and can be used to detect the antiserum of HV virus.</p>
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Base de dados:
WPRIM
Assunto principal:
Células Vero
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Expressão Gênica
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Chlorocebus aethiops
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Proteínas do Envelope Viral
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Baculoviridae
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Orthohantavírus
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Vetores Genéticos
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Genética
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Metabolismo
Limite:
Animals
Idioma:
Zh
Revista:
Chinese Journal of Experimental and Clinical Virology
Ano de publicação:
2011
Tipo de documento:
Article