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Fig. 3 Rescue of the mys mutant phenotype by transgenesis. (A) The structure of the Tol2–mys construct. mys cDNA (mys) was cloned between the Xenopus EF1α promoter (EF1α-p) and the SV40 polyA signal (poly A). Thick black lines indicate the Tol2 sequence. (B) A transposon-donor plasmid containing Tol2–mys and the transposase mRNA were co-injected into fertilized eggs produced from mys homozygous parents. The injected fish were crossed with mys homozygous fish and transgenic fish were identified in the progeny. A female homozygous for the mys mutation (mys/mys) and carrying a single copy insertion of Tol2–mys was crossed with a wild type male, and the progeny were analyzed for the phenotype. (C, D) Dorsal views of embryos at the 6-somite stage. (C) Embryos from homozygous (mys/mys) female fish showed the obscure somite boundary phenotype. (D) Embryos from mys/mys; Tg(Tol2–mys) female fish showed distinct somite boundaries.
Reprinted from Developmental Biology, 316(2), Kotani, T., and Kawakami, K., misty somites, a maternal effect gene identified by transposon-mediated insertional mutagenesis in zebrafish that is essential for the somite boundary maintenance, 383-396, Copyright (2008) with permission from Elsevier. Full text @ Dev. Biol.