Grabher C, Henrich T, Sasado T, Arenz A, Wittbrodt J, Furutani-Seiki M
Developmental Biology Programme, EMBL, Meyerhofstr. 1, D-69012 Heidelberg, Germany.
Gene. 2003 Dec 11;322:57-66. doi: 10.1016/j.gene.2003.09.009.
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.
我们测试了睡美人转座元件高效地将转基因插入青鳉（Oryzias latipes）基因组的能力，青鳉是脊椎动物发育的重要模型系统。我们发现SB转座子能有效地介导报告基因整合到鱼类生殖系中。在初步实验中，我们建立了174个转基因品系，转基因效率为32%。转基因可稳定地传递给后代并在后代中表达。有趣的是，转基因品系以12%（21/174）的比例呈现出具有时间和空间特异性的新型表达模式，这可能是由于增强和沉默位置效应共同作用的结果。此外，注射的鱼胚胎中依赖启动子的绿色荧光蛋白表达与生殖系传递紧密相关，便于轻松筛选出奠基鱼。因此，SB转座子/转座酶系统总体上为转基因提供了一种高效工具，尤其为产生新型报告基因表达模式提供了高效工具。
