YAN Yan,etc.,al. Construction of reassortant H1N1 influenza A virus by eight-plasmid system. Journal of Northwest A & F University
To establish the reverse genetic system for influenza virus,Hemagglutinin(HA) and Neuraminidase(NA) gene of A/shanghai/7/99 subtype H1N1 were amplified by RT-PCR,then were cloned into the vector pGEM-T and sequenced.The positive clone was digested by BsmB Ⅰ and Bsa Ⅰ,seperately,then inserted into the bi-directional expression vector pAD3000 to construct recombinant plasmids:pAD-HA and pAD-NA.They were transfected into COS-1 cells in combination with plasmids incorporating the other 7 genes of A/PR/8/34 resp...
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