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. 2013 Apr 24;8(4):e62690. doi: 10.1371/journal.pone.0062690

Figure 4. Testing elements for (A) promoter and (B) terminator activities in S2 cells.

Figure 4

In the structural scheme of the scs insulator (PvuII–PvuII fragment), the Zw5 site is shown as a white rectangle; identified polyadenylation sites (PAS), as black circles with “+” sign indicating their direct orientation. Restriction sites HpaI–NdeI indicate the boundaries of the element used in the terminator assay. In the reductive schemes of transgenic constructs, the ORFs encoding Renilla (Rluc) and firefly (Fluc) luciferases are shown as white and black boxes with arrows indicating the direction of transcription; the white rectangle marked “term” is the SV40 terminator. The bicistronic plasmid also contained the actin promoter and rpr IRES (rectangles with corresponding marks). Thick downward arrows indicate insertion sites for the hsp70 promoter (Hsp70 pr), scs (scs dir, scs rev), and scs' (scs' dir, scs' rev) in the promoter assay and for late SV40 (termSV40) and scs PASs (scs(polyA)dir, scs(polyA)rev) in the terminator assay. The reporter system used in the assays is based on measurement of Fluc versus Rluc activity. The Fluc/Rluc ratios for the test constructs are shown in histograms. Error bars show standard deviations (n = 3).