FIG. 3.

Synthesis and virion localization of the G7L protein. (A) Temporal synthesis of the G7L protein. BS-C-1 cells were mock infected for 8 h (lane U) or infected with VV at a multiplicity of infection of 10 in the absence or presence of AraC and harvested between 0 and 24 h postinfection (hpi). Proteins from total cell extracts were resolved by electrophoresis on an SDS-4 to 20% gradient polyacrylamide gel and analyzed by Western blotting using rabbit G7L peptide antiserum. Proteins were detected by chemiluminescence. The bands corresponding to the full-length and cleaved forms of the G7L protein are indicated on the right. The positions of migrations and molecular masses of marker proteins are indicated on the left. (B) Association of the G7L protein with purified virions. Sucrose gradient-purified VV was incubated in buffer containing 1% NP-40 with or without 50 mM dithiothreitol. After centrifugation, the soluble (S lanes) and insoluble (P lanes) fractions were analyzed by Western blotting using G7L or A14L peptide antiserum. Bands corresponding to the A14L protein or to the full-length and cleaved forms of the G7L protein are indicated on the right. The numbers on the left correspond to the molecular masses of the marker proteins. DTT, dithiothreitol.