Xenbase Image ID: 81565

Figure 4. BAC constructs replicate more efficiently in developing embryos than smaller plasmids. (a) Schematic showing pax6 dual reporter BAC injected into embryos (40 pg). DNA gel image shows results from PCR assay amplifying gfp3 at the indicated stages. (b) Schematic showing plasmid reporter injected into embryos for PCR replication assay. DNA gel images show PCR results from two different DNA amounts injected (40 pg and 4 pg, as indicated). (c) pax6 promoter-GFP in BAC vector construct injected into embryos for PCR assay. Gel shows PCR results from embryos collected at indicated stages. (d) Graph shows results of quantification of gel bands from three independent experiments performed for each construct, and though these experiments were done independently using slightly different PCR conditions, they are shown together in Figure 4d to illustrate the difference in patterns in the two experiments. Error bars show standard deviations, and asterisks indicate P values <0.05 for transgene amplification as compared to 1 hpf at stages 9 (P = 0.016), 25 (P = 0.037), and 35 (P = 0.016). (e) Consistent expression of transgene is observed in pax6 BAC injected embryos. (f) Ectopic, mosaic, and inconsistent expression observed in plasmid injected embryos. Reproduced with permission of the Publisher, John Wiley & Sons Image published in: Fish MB et al. (2012) Copyright © 2012. Image reproduced with permission of the Publisher, John Wiley & Sons. Larger Image Printer Friendly View

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