Fig. 1.
Strategy for cloning the Torso (Tor), Tsl and Trk open reading frames into the multicistronic expression vectors pAc5-STABLE2-Neo and pAc5-STABLE1-Neo. (Top row) Schematic diagram of the multicistronic segment of pAc5-STABLE2-Neo (Gonzalez et al., 2011), which contains the Actin5C promoter, and mCherry, GFP and NeoR genes, is shown. Restriction sites and the positions of the dT2A and T2A autocleavage sites (black and gray boxes, respectively) are shown. (Rows 2-5) Plasmid derivatives in which DNA fragments encoding wild-type Tor/Tor[4021], Tsl and/or Trk have been introduced. (Row 6) Schematic diagram of the bicistronic region of pAc5-STABLE1-Neo (Gonzalez et al., 2011), which carries the Actin5C promoter and GFP and NeoR open reading frames, with restriction sites indicated. (Row 7) The plasmid encoding a Tor-GFP fusion protein (pAc5-STABLE1-Tor-GFP-Neo), with the cloning sites that were used for the substitution displayed as well as other restriction sites that remain. pAc5-STABLE1-Toll-GFP-Neo, which encodes a Toll-GFP fusion protein, is shown in the last row. B, BamHI; E1, EcoRI; EV, EcoRV; H, HindIII; K, KpnI; Nh, NheI; No, NotI; Xb, XbaI; Xh, XhoI.