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. 2011 Jun;85(11):5350–5362. doi: 10.1128/JVI.00035-11

Fig. 2.

Fig. 2.

VP80 copurifies with nucleocapsids of both BV and ODV virions and is associated with one end of the nucleocapsid. (A) Ac-Δvp80-Flag:vp80 BV virions were separated into nucleocapsid (Nc) and envelope (Env) fractions. The Flag-VP80 protein was detected on immunoblots with anti-Flag antibody (upper panel). Correct separation into Nc and Env fractions was controlled with anti-VP39 and anti-GP64 antibodies (bottom panels). (B) The ODV virions produced in Sf9 cells coinfected with the Ac-Δvp80-Flag:vp80 (MOI = 25) and AcMNPV strain E2 (MOI = 5) viruses were separated similarly into Nc and Env fractions and immunoblotted with anti-Flag antibody (upper panel). Proper fractionation was controlled using anti-PIF-1 antiserum (bottom panel). Equal amounts of protein (10 μg) were loaded in the lanes. (C) Immunoelectron microscopy of BV nucleocapsids derived from either the Ac-Δvp80-Flag-vp80 (a) or Ac-Δvp80-egfp:vp80 (b) genotype. (c, d) Nucleocapsids purified from the Ac-wt genotype-propagated BVs were used as a negative control. Purified nucleocapsids were adsorbed to EM grids and immunostained with mouse anti-Flag (a, c) or anti-EGFP (b, d) antibody, followed by goat anti-mouse immunoglobulin conjugated with 5-nm gold particles. The black arrowheads point to specific localizations of 5-nm gold particles. Bars represent 200 nm.