PURIFICATION AND ANALYSIS CATALYTIC FUNCTIONS OF RECOMBINANT INFLUENZA VIRAL POLYMERASES

Polymerase of influenza virus is made up three subunits PB1, PB2, and PA, which are involved in viral genome transcription replication. Purification sufficient amounts polymerase essential to understand the catalytic function polymerase. In this study, we generated a expression system human embryonic kidney 293T cells (293T cells). The cDNAs for RNA segments 1, 2, 3, encode PA proteins, respectively, were integrated into mammalian plasmids pCAGGS simultaneously express all proteins cells. We purified recombinant polymerases A/PR/8/34 (H1N1) (PR8) avian A/Turkey/England/1969 (H3N2) (TE) using anti-FLAG M2 affinity resin. After confirming trimeric complexes, enzymatic properties characterized, including model binding, vitro assays, cap-snatching activity upon addition cap1-70 mer vRNA as substrate. Taken together conclude that suitable amount isolation functional with 95% purity.