Figure 1

Construction of a myf5:GFP bacterial artificial chromosome (BAC) and deletion constructs for germ-line transmission in zebrafish. (A) Strategy for constructing a myf5 BAC clone containing the green fluorescent protein (GFP) reporter. (Top) The genomic organization of the myf5 that contains 3 exons (E1, E2, and E3) and 2 introns (I1 and I2). (Bottom) The resulting p(myf5(80K):GFP) clone contains the myf5 upstream 80-kb regions fused with the GFP reporter gene. The primers ZMFP-117F, GFP-R, Kan-F, and ZMF-1000R were used to check recombinants. (B) Deletion constructs used in this study. Plasmid pZMYP-2456E was described by Wang et al. [13]. Thick lines and crossed boxes represented plasmid vectors and myf5 promoters, respectively. Numbers above the boxes indicate the nucleotide positions relative to the transcription start site of zebrafish myf5. GFP, green fluorescent protein; ITR, inverted terminal repeats of adeno-associated virus; SVpA, polyadenylation signal of SV40.