Skip to main content
. 2008 Oct 31;36(21):6907–6917. doi: 10.1093/nar/gkn793

Figure 3.

Figure 3.

Verification of SSA in transiently transfected S2 cells. (A) Constructs for transfection are depicted: enhancer/promoter-containing mtnB copy (MtnBgen) and enhancer/promoter-less mtnb-EGFP fusion copy (MtnBgen-EGFP). For the positive control construct for intramolecular SSA (MtnBgen-gen), see Figure 1B. Constructs indicated in the bar diagram were always cotransfected with a plasmid expressing the endonuclease I-SceI. Green fluorescence as a measure for SSA was calculated as described in Materials and Methods section. Error bars represent standard deviations of three experiments. (B) Isolation of SSA products from S2 cells. EGFP positive cells were first sorted, and plasmid DNA was isolated. PCR to detect SSA products was performed. The indicated tester constructs are depicted inFigure 1B. PCR products of plasmid DNA from EGFP positive S2 cells after DSB induction (S2) and from E. coli without induction of DSB (Ec) are shown. Expected PCR bands for SSA products are 431 bp and 370 bp (with and without the intron). The three marker bands shown indicate the position of 300 bp, 400 bp and 500 bp.