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Analysis of TAP-Purified Influenza Polymerases from Nanchang E627 and E627K Mutants

This study presents SDS-PAGE data illustrating the TAP-purified influenza polymerases from Nanchang E627 and the mutant E627K. The analysis indicates an underestimation of the concentration for the mutant polymerase, quantified at 1.5 times that of the E627 polymerase. The purification process, reliant on a tagged PA, may lead to varying amounts of excess PA. It's important to note that only the trimeric form of the polymerase is enzymatically active, which necessitated normalization based on PB1 and PB2 concentrations rather than PA.

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Analysis of TAP-Purified Influenza Polymerases from Nanchang E627 and E627K Mutants

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  1. Supplementary Data 4 Nan E627 Nan E627K PB1 PA PB2 0.88 :PB1 1 :PB2 0.91 1 Supplementary Data 4. SDS-PAGE showing TAP-purified influenza polymerases: Nanchang E627 (5μl) and Nanchang E627K (10μl). In approximation, we underestimated the concentration of the mutant polymerase to 1.5X of the E627 polymerase. Note: Since the purification relies on a tagged-PA, excess PA can be pulled depending on the purification. However, only a trimer is active, therefore we normalized for PB1 and PB2 concentrations rather than PA.

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