Establishment of reverse genetics system for class I NDV08-004 strain / 病毒学报
Chinese Journal of Virology
; (6): 496-500, 2012.
Article
em Zh
| WPRIM
| ID: wpr-340017
Biblioteca responsável:
WPRO
ABSTRACT
Based on the genomic sequence of NDV08-004 strain (GenBank accession number FJ794269), seven pairs of primers were designed to amplify the genomic fragments by RT-PCR and cloned into pGEM-Teasy vector. The fragments (named A to G) were sub-cloned into transcription vector pOLTV5 according to the universal RE site and the plasmid named NDV08-004-pO which contained the full length cDNA of NDV08-004 strain was constructed. Three helper plasmids (pCI-NP, pCI-P and pCI-L) together with NDV08-004-pO were co-transfected into BSR T7/5 cells, and the transfection supernatant was inoculated into SPF embryonated eggs to rescue the virus. The virus was rescued successfully and identified by HA and RT-PCR and sequencing. The rescue system constructed in this study provided a good foundation for the further related research.
Texto completo:
1
Índice:
WPRIM
Assunto principal:
Plasmídeos
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Virologia
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Vírus da Doença de Newcastle
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Dados de Sequência Molecular
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Sequência de Bases
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Genética Reversa
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Vetores Genéticos
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Genética
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Métodos
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Doença de Newcastle
Limite:
Animals
Idioma:
Zh
Revista:
Chinese Journal of Virology
Ano de publicação:
2012
Tipo de documento:
Article