FIG. 2.

Specific transcription reactions reconstituted with partially purified VLTF-X or virion extracts (VEs). Shown is an autoradiogram of a 4% denaturing polyacrylamide gel in which the radiolabeled RNA products of specific transcription reactions were separated. In lanes 1 to 3 all reaction mixtures contained 3 μl of G8R protein, 2 μl of A1L protein, 4 μl of A2L-GST fusion protein, and 3 μl of the viral RNA polymerase purified from infected HeLa cells (cpol). In addition, the reaction mixture for lane 2 contained 5 μl of hydroxylapatite-purified VLTF-X [VLTF-X (HAP)] and that for lane 3 contained 5 μl of phosphocellulose II-purified VLTF-X [VLTF-X (Pcell II)]. In lanes 4 to 7, 3 μl of G8R protein, 3 μl of A1L protein, and 2.5 μl of A2L protein were present in all reaction mixtures. In addition, the reaction mixtures for lanes 4, 5, and 7 contained 3 μl of viral RNA polymerase purified from infected HeLa cells. Lanes 5 and 6 additionally contained 2.5 μl of a soluble extract (VE) made from sucrose gradient-purified vaccinia virions that was capable of transcribing vaccinia virus early genes (19). Lane 7 additionally contained 5 μl of the flowthrough of two successive DEAE-cellulose columns (DEAE FT) used to purify VETF from the crude virion extracts (19).