Influenza A Virus (IAV) subtypes, A/England/195/2009 (pH1N1) and the A/HK/1174/99 (H3N2), were obtained as described recently (32 (link), 33 (link)). H1N1 and H3N2 pseudotyped viral particles were produced using the plasmids obtained from Addgene (32 (link), 33 (link)). pcDNA3.1-swineH1-flag (H1 from swine H1N1 A/California/04/09) (Invitrogen), pcDNA3.1-swine N1-flag (N1 from swine H1N1 A/California/04/09), and pcDNA H3 (from A/Denmark/70/03 (H3N2) (Invitrogen) were purchased commercially. Plasmid pI.18-N2 [N2 from A/Texas/50/2012/(H3N2)] was provided by Dr Nigel Temperton (University of Kent). Anti-influenza viral antibodies and recombinant viral proteins were obtained from BEI Resources, National Institutes of Health, USA (32 (link), 33 (link)).
Pcdna h3
Manufactured by Thermo Fisher Scientific
The pcDNA H3 is a plasmid vector commonly used for the expression of proteins in mammalian cell lines. It contains a human cytomegalovirus (CMV) promoter, which drives the expression of the gene of interest. The plasmid also includes a neomycin resistance gene, which allows for selection of cells that have been successfully transfected with the vector.
Automatically generated - may contain errors
2 protocols using pcdna h3
1
Influenza Virus Subtypes Propagation
2
Generating Influenza Pseudotyped Viral Particles
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The IAV subtypes used in this study, including the pH1N1 (A/England/2009) and H3N2 (A/HK/1999), were provided by Wendy Barclay (Imperial College, London) and Leo Poon (University of Hong Kong), respectively. The plasmids used for the production of H1N1 and H3N2 pseudo-typed viral particles were obtained from Addgene; pHIV-Luciferase (Addgene plasmid #21375); psPAX2 (Addgene plasmid # 12260); and Vesicular Stomatitis Virus (VSV-G) (Addgene plasmid #8454). pcDNA3.1-swineH1-flag (H1 from swine H1N1 A/California/04/09) (Invitrogen), pcDNA3.1-swine N1-flag (N1 from swine H1N1 A/California/04/09), and pCDNA H3 (from A/Denmark/70/03 (H3N2) (Invitrogen) were obtained commercially. pI.18-N2 [N2 from A/Texas/50/2012/(H3N2)] plasmid was a gift from Nigel Temperton (University of Kent). Anti-influenza antibodies used were obtained from BEI Resources, NIAID, NIH, USA, and used as previously described (38 (link)); these include polyclonal anti-influenza Virus H3 HA, A/Hong Kong/1/1968 and monoclonal anti-influenza virus H1 HA, A/California /04/2009.
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