FIG. 1.

Construction, rescue, and characterization of mutant measles virus. (A) Schematic representation of the measles virus genome. The mutant H gene was replaced in the full-length plasmid coding for the measles genome by use of PacI and SpeI restriction sites. (B) Rescue of the recombinant virus was documented by the presence of infection in Vero cells that expressed high levels of GFP without significant cell-to-cell fusion. (C) One-step growth curve of released and cell-associated virus for MVeGFP and MV-I98A after infection of Vero cells at a multiplicity of infection (MOI) of 1. (D) Western blot analysis of MVeGFP and MV-I98A by use of antibodies against measles N, H, and F proteins. The amount of purified virus loaded ranged from 5 × 10³ to 30 × 10³ 50% tissue culture infectious doses (TCID₅₀). (E) Coimmunoprecipitation of MV H-F complexes. Vero cell lysates (CL) collected after infection with MVeGFP or MV-I98A were pulled down with protein G agarose beads conjugated to mouse anti-MV-H IgG, and MV-F protein was then detected by Western blot analysis using rabbit anti-MV-F primary antibody. MVeGFP virions (V) were loaded at a TCID₅₀ of 5 × 10⁴. Wt, wild type.