FIG 5.

Analysis of BV production. (A) Extracellular BV titers from Sf9 cells separately transfected with Ac-MK, Ac11KO, and Ac11Re were determined at 24 and 72 hpt by TCID₅₀ endpoint dilution. Each point represents the average titer derived from three independent TCID₅₀ assays. The error bars represent standard deviations. (B) The virus titer, regardless of virion infectivity, of the supernatants of Sf9 cells transfected with Ac-MK, Ac11KO, or Ac11Re was determined by qPCR analysis at the designated time points. A stock of wild-type Ac-MK (4.25 × 10⁸ PFU/ml) whose titer was previously determined by endpoint dilution was serially diluted and used to develop a standard curve (shown in the inset). (C) Western blot analysis of BV isolated from Sf9 cells transfected with Ac-MK, Ac11KO, or Ac11Re at 72 hpt. The blots were probed with a monoclonal antibody specific for the VP39 nucleocapsid protein. (D) Detection of viral DNA in Sf9 cells infected with serially passaged Ac11KO. The passage numbers (P2 to P5) are indicated for Sf9 cells infected with serially passaged Ac11KO.