Mutations in the CP can affect the inhibition of reporter protein production. (A) Expression of the mutant BMV CPs in N. benthamiana. The mutant CPs were all constructed by site-directed mutagenesis and verified to have only the intended mutation. The amount of CP produced in N. benthamiana was examined by a Western blot probed with antiserum to the BMV CP. The small subunit of the Rubisco protein was used as a loading control. (B) A survey of the effects of the mutant CPs on the inhibition of GFP production from the 2GFP2 reporter. N. benthamiana plants were infiltrated to express 2GFP2 (OD595 of 0.2) and either a culture containing the empty vector, the wild-type CP, or a select mutant CP. The leaves were macerated at 48 hpi and subjected to Western blot analysis detecting GFP. (C) Further examination of the effects of D139A on GFP accumulation from construct 2GFP2. The densities of the cultures expressing the WT CP or the D139A mutant are shown above the Western blot image.
