Figure 1

RNP assaying by Fluc and Gluc systems after reconstitution of influenza A virus polymerase complex. (A) Polymerase activity assayed by a viral UTR-driven Gluc reporter gene. (B) Detection and quantification of RNP activity by the Gluc or Fluc reporter systems after transfection. 293T cells were transfected with plasmids encoding the PB2, PB1, PA, and NP genes of A/Quail/HK/G1/97 (H9N2) plus reporter plasmids polI-Gluc or pYH-Fluc. Supernatants (cell-free culture medium) and cell lysates were assayed for luciferase activity using appropriate substrates by a luminometer. (C) Kinetics of G1 RNP activity assayed using the Gluc reporter system after virus infection. A549 cells were transfected with plasmid polI-Gluc and infected at 6 h post-transfection with influenza virus G1 at a MOI of 1. Luciferase activity in supernatants was assayed at 0, 3, 6, 9, 12, and 24 h post-G1 (H9N2) infection and expressed as RLU per well. Data are expressed as standard errors of the mean (SEM) from three independent experiments performed in triplicate.