High growth influenza virus
An influenza virus, influenza B virus technology, applied in the field of use, can solve problems such as inability to guarantee protection
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Embodiment 1
[0296] Purpose:
[0297] The 6:2B / Thuringia / 02 / 06:B / Murmansk / 3 / 2010delNS1 point mutant was tested for growth in a growth curve assay to identify mutations that resulted in improved virus growth.
[0298] method
[0299] Generation of 6:2 recombinant viruses with specific amino acid changes
[0300] To generate amino acid changes in internal genes, point mutations were performed in plasmids containing the gene of interest by subjecting each plasmid to site-directed mutagenesis using the QuikChange Lightning Site-Directed Mutagenesis Kit (Agilent, Santa Clara, CA). Generation of 6:2 reassortant viruses by reverse genetics. Six pHW2000 derivatives containing fragments PB2, PB1, PA, NP, M, ΔNS1 derived from B / Thuringia / 02 / 06 (a B / Jiangsu / 10 / 03-like virus from the B Yamagata lineage) (plasmid) and a protein expression plasmid encoding influenza A PR8 NS1 (pCAGGS-NS1(SAM)) co-transfected into Vero cells with a pHW2000 derivative plasmid containing the HA and NA genes from B / Murma...
Embodiment 2
[0330] Purpose:
[0331] The 6:2B / Thuringia / 02 / 06:B / Phuket / 3073 / 2013delNS point mutant was tested in a growth curve assay to identify mutations that lead to improved virus growth.
[0332] method:
[0333] Generation of 6:2 recombinant viruses with specific amino acid changes
[0334] To generate amino acid changes in internal genes, point mutations were performed in plasmids containing the gene of interest by subjecting each plasmid to site-directed mutagenesis using the QuikChangeLightning Site-Directed Mutagenesis Kit (Agilent, Santa Clara, CA). Generation of 6:2 reassortant viruses by reverse genetics. Six pHW2000 derivatives containing fragments PB2, PB1, PA, NP, M, ΔNS1 derived from B / Thuringia / 02 / 06 (a B / Jiangsu / 10 / 03-like virus from the B Yamagata lineage) (plasmid) and a protein expression plasmid encoding influenza A PR8 NS1 (pCAGGS-NS1(SAM)) were co-transfected into Vero cells with a pHW2000 derivative plasmid containing the HA and NA genes from B / Puji / 3073 / 2013 ...
Embodiment 3
[0361] Purpose:
[0362] The 6:2 A / IVR-116:A / Hong Kong / 4801 / 2014 delNS1 point mutant was tested for growth in a growth curve assay to identify mutations that lead to improved virus growth.
[0363] method:
[0364] Generation of 6:2 recombinant viruses with specific amino acid changes
[0365] To generate amino acid changes in internal genes, point mutations were performed in plasmids containing the gene of interest by subjecting each plasmid to site-directed mutagenesis using the QuikChange Lightning Site-Directed Mutagenesis Kit (Agilent, Santa Clara, CA). Generation of 6:2 reassortant viruses by reverse genetics. A laboratory virus containing the PB2, PA, NP, M, and NS genes from A / Puerto Rico / 08 / 1934 and PB1 from A / Texas / 1 / 1977, derived from A / IVR-116, will be Six pHW2000 derivatives (plasmids) of fragments PB2, PB1, PA, NP, M, and ΔNS1 of strain A virus) and a protein expression plasmid (pCAGGS-NS1 (SAM)) encoding influenza A PR8 NS1 combined with those from A / HK / 480...
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