The Gurdon Institute and Department of Genetics, University of Cambridge, Cambridge, United Kingdom.
PLoS One. 2011;6(5):e20082. doi: 10.1371/journal.pone.0020082. Epub 2011 May 26.
Here we describe a toolkit for the production of fluorescently tagged proteins in the C. elegans germline and early embryo using Mos1-mediated single copy insertion (MosSCI) transformation. We have generated promoter and 3'UTR fusions to sequences of different fluorescent proteins yielding constructs for germline expression that are compatible with MosSCI MultiSite Gateway vectors. These vectors allow tagged transgene constructs to be inserted as single copies into known sites in the C. elegans genome using MosSCI. We also show that two C. elegans heat shock promoters (Phsp-16.2 and Phsp-16.41) can be used to induce transgene expression in the germline when inserted via MosSCI transformation. This flexible set of new vectors, available to the research community in a plasmid repository, should facilitate research focused on the C. elegans germline and early embryo.
在这里，我们描述了一种使用 Mos1 介导的单拷贝插入（MosSCI）转化在秀丽隐杆线虫生殖系和早期胚胎中产生荧光标记蛋白的工具包。我们已经生成了与不同荧光蛋白序列的启动子和 3'UTR 融合，产生了适合 MosSCI MultiSite Gateway 载体的生殖系表达构建体。这些载体允许标记的转基因构建体通过 MosSCI 以单拷贝的形式插入秀丽隐杆线虫基因组中的已知位点。我们还表明，当通过 MosSCI 转化插入时，两个秀丽隐杆线虫热休克启动子（Phsp-16.2 和 Phsp-16.41）可以用于诱导生殖系中转基因的表达。这套灵活的新载体以质粒库的形式提供给研究社区，应该有助于专注于秀丽隐杆线虫生殖系和早期胚胎的研究。
