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. Author manuscript; available in PMC: 2009 Feb 14.
Published in final edited form as: Cell Host Microbe. 2008 Aug 14;4(2):111–122. doi: 10.1016/j.chom.2008.06.007

Figure 5. Defective assembly of mutant polymerase into vRNP is overcome during late stages of infection.

Figure 5

(A) Recombinant virus was produced in 293T cells transfected with pTM-pol I-WSN-All and expression vectors for NP, PB1, PA, PB2, and WT or K627E PB2-TAP. PB2 and PB2-TAP were detected by western blot of cell lysate or cell-free virions with anti-PB2 serum. (B) PB2-TAP was precipitated with rabbit IgG-agarose from virus-producing cell lysates. Co-precipitating NP was detected by anti-NP western blot. (C) Purified virus was lysed and separated on a linear 30–60% glycerol gradient. Where indicated, 1 % deoxycholate (DOC) was present during viral lysis. Fractions were collected and PB2 and NP were detected by western blot. Non-specific bands at the top of the gradient are indicated (*).