RRC ID 87776
著者 Grabher C, Henrich T, Sasado T, Arenz A, Wittbrodt J, Furutani-Seiki M.
タイトル Transposon-mediated enhancer trapping in medaka.
ジャーナル Gene
Abstract We tested the Sleeping Beauty transposable element for its ability to efficiently insert transgenes into the genome of medaka (Oryzias latipes), an important model system for vertebrate development. We show that the SB transposon efficiently mediates integration of a reporter gene into the fish germ line. In pilot experiments, we established 174 transgenic lines with a transgenesis efficiency of 32%. Transgenes are stably transmitted to, and expressed in, subsequent generations. Interestingly, the transgenic lines show novel expression patterns with temporal and spatial specificity at a rate of 12% (21/174), likely due to both, enhancing and silencing position effects. Furthermore, promoter-dependent GFP expression in injected fish embryos is tightly correlated with germ line transmission, facilitating easy selection of founder fish. Thus, the SB transposon/transposase system provides a highly efficient tool for transgenesis in general and for the generation of novel reporter gene expression patterns in particular.
巻・号 322
ページ 57-66
公開日 2003-12-11
DOI 10.1016/j.gene.2003.09.009
PII S0378111903008758
PMID 14644497
MeSH Animals Animals, Genetically Modified Base Sequence Blotting, Southern DNA / chemistry DNA / genetics DNA Transposable Elements / genetics* Embryo, Nonmammalian / metabolism Gene Expression Regulation, Developmental Green Fluorescent Proteins Luminescent Proteins / genetics Luminescent Proteins / metabolism Microinjections Microscopy, Fluorescence Molecular Sequence Data Mutagenesis, Insertional / methods* Oryzias / embryology Oryzias / genetics* Plasmids / administration & dosage Plasmids / genetics Recombinant Fusion Proteins / administration & dosage Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / metabolism Sequence Analysis, DNA
IF 2.984
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