Fig. 1

Description of PhOTO Vector and PhOTO Transgenics.

(A) Depicted is a schematic of the general PhOTO vector. The major components of the vector, including the promoter, the TaV 2A sequence, the Tol2 transposable elements, and the protein locations are indicated on the outside of the plasmid circle. Each vector was designed such that each of the components (e.g. the promoter, the FPs, etc.) may be easily switched out for alternate protein fusions, etc. using the listed restriction enzymes (inside the circle) and an appropriate subcloning procedure. Note that the sizes of the blocked regions indicating coding sequences are not to scale. (B) Representative heterozygous PhOTO-N expression in an 18–19 hour post fertilization F1 transgenic zebrafish embryo. The top left panel of (B) depicts memb-Cerulean (blue); the top right panel depicts unconverted H2B-Dendra2 (green); the bottom left panel depicts photoconverted H2B-Dendra2 (red) in 4 somites, the tip of the tail, a subset of cells in the eye, and a subset of cells atop the yolk; and the bottom right panel depicts a merged image of all three colors. Scale bar is 100μm.