Fig. 4

Test of pDestTol2CG destination vector. Embryos were injected at the one-cell stage with 25 pg of DNA for pDestTol2CG; hsp70: mCherryCAAX-polyA, either with or without 25 pg of transposase RNA. A: Schematic of expression construct and graph showing fraction of embryos with a green heart at 30 hours postfertilization (hpf). Arrows in schematic indicate direction of transcription. Coinjection of transposase RNA greatly increased the fraction of embryos with green hearts. B-E: Further analysis of embryos selected at 30 hpf for green hearts. B-B′″: Embryos at 48 hpf, after heat shock for 1 hr at 30 hpf. C-C<′″: Embryos at 48 hpf, without heat shock. No red fluorescence is detectable at this magnification (C″).D-D′″: Close up of two heat-shocked embryos, shown at 48 hpf. Note bright red fluorescence (D″) and lack of ectopic green fluorescence (D′).E-E′″: Close up of two non-heat-shocked embryos, shown at 48 hpf. Note that there is no mCherry fluorescence in the heart. Dim, diffuse red fluorescence appears in one embryo (E″, arrowhead), indicating leaky expression from the hsp70 promoter.