The rescue of H1N1 subtype swine influenza virus / 生物工程学报
Chinese Journal of Biotechnology
; (12): 857-861, 2008.
Article
en Zh
| WPRIM
| ID: wpr-342826
Biblioteca responsable:
WPRO
ABSTRACT
The swine influenza virus (SIV) strain A/Swine/TianJin/01/2004(H1N1) (A/S/TJ/04) was rescued successfully by an eight-plasmid rescue system. The cDNAs of SIV 8 gene segments were synthesized by RT-PCR and cloned into the RNA polymerase I/II bidirection expression vector PHW2000 independently, resulting in 8 recombinant plasmids. The 8 recombinant plasmids were cotransfected into COS-1 cell, 30 h later TPCK-trypsin was added to 0.5 microg/mL. The COS-1 cell and supernatant were harvested 48 h after cotransfection and were inoculated into the allantoic cavity of 9-day-old specific-pathogen free (SPF) chicken eggs. The allantoic fluid of dead eggs was harvested and passaged 3 generations in SPF chicken eggs to get infective virus. The successful rescue of A/S/TJ/04 SIV was identified by hemagglutination assay, hemagglutination inhibition assay, sequence analysis and electron microscope observation. The successful rescue of SIV built a platform for the research of the relationship between genome structure and function of SIV, the mechanisms of trans-species transmission of influenza virus and for the generation of new SIV as vaccine.
Texto completo:
1
Base de datos:
WPRIM
Asunto principal:
Fisiología
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Plásmidos
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Recombinación Genética
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Porcinos
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Replicación Viral
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ARN Polimerasa I
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ARN Polimerasa II
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Transfección
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Chlorocebus aethiops
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Pollos
Límite:
Animals
Idioma:
Zh
Revista:
Chinese Journal of Biotechnology
Año:
2008
Tipo del documento:
Article