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Fig. 1 Isolation of transgenic T2/OncZ concatemer lines.
(A) T2/OncZ transposon vector and RFP reporter used to isolate concatemers. IRL and IRR, left and right transposon inverted repeats; SA, splice acceptor from intron 1 of ß-actin gene; MSCV 52 LTR, murine stem cell virus 52 long terminal repeat; β-actin, β-actin promoter minus splice acceptor at 3′ end; SD, splice donor; En2-SA, splice acceptor from mouse engrailed 2 gene; pA, SV40 polyadenylation sequence; AFP, ocean pout antifreeze protein 3′ UTR; black bar represents probe used on genomic Southerns shown in panel C. Grey box represents probe used on genomic Southerns shown in panel D. Linear DNA fragments of T2/OncZ and the β-actin:RFP reporter gene were mixed and co-injected into 1-cell zebrafish embryos. (B) Adult F0 founders were outcrossed to wild type and transgenic F1 embryos identified by ubiquitious RFP fluorescence. (C) Genomic Southern blots to estimate transposon copy number in Tg(T2/OncZ, β-actin:RFP) concatemer lines 1–7. DNA was isolated from F2 generation heterozygous adults. (D) Genomic Southern blot of DNA isolated from F3 generation heterozygous Tg(T2/OncZ, β-actin:RFP)is7 and Tg(T2/OncZ, β-actin:RFP)is6 adults. Plasmid pT2/OncZ was loaded as reference in copy # control lanes.