Genetic linkage of zebrafish ocular tumor model in transgenic line Tg(flk1:RFP)is18.
(A) Endothelial-specific membrane targeted RFP expression in live 3 dpf zebrafish larva from transgenic line Tg(flk1:RFP)is18. (B) Endothelial-specific membrane targeted RFP expression and nuclear eGFP expression in live 3 dpf Tg(flk1:RFP)is18; Tg(fli1:eGFP)y1 zebrafish larva. (C) Gross morphology of ocular tumors in heterozygous is18/+ adults at 5 months and 1 year of age. (D) Pedigree of one family showing trans-generational linkage analysis of ocular tumor phenotype with inheritance of the Tol2<flk1:RFP> transgene in heterozygous is18/+ individuals. Two families, A and B, were established from the F4 generation. Marked individuals were used in subsequent STR/STS linkage (L/S) and Southern blot analyses (#). (E) Diagram of endothelial-specific RFP reporter construct pTol2<flk1:RFP-CAAX> with positions of EcoRI and SacI sites. Genomic Southern blot hybridized with RFP probe detects 1.3 kb EcoRI -SacI fragment (arrows) of the Tol2<flk1:RFP-CAAX> transgene. The high copy-number Tol2<flk1:RFP> transgene array is transmitted specifically to RFP expressing, tumor positive Tg(flk1:RFP)is18 progeny through the F6 generation.