Figure 3.

(A) Histidine-tagged dTFIIE-L and dTFIIE-S were expressed individually in bacteria, extracted, and purified as described. The two subunits were combined and renatured by stepwise dialysis. Soluble dTFIIE complex was subjected to SDS/PAGE and stained with Coomassie blue (lane 2). The mobility of the large (IIE-L) and small (IIE-S) subunits are indicated. The identity of the band directly above IIE-S is unknown and most likely a bacterial contaminant. (B) Western blot analyses of bacterially expressed untagged dTFIIE-L (lane 2), dTFIIE-S (lane 4), and Drosophila embryo NE (lanes 1 and 3). Blots were probed with antibodies raised against recombinant dTFIIE-L (lanes 1 and 2) and dTFIIE-S (lanes 3 and 4).