MosSCI and gateway compatible plasmid toolkit for constitutive and inducible expression of transgenes in the C. elegans germline.

MosSCI and gateway compatible plasmid toolkit for constitutive and inducible expression of transgenes in the C. elegans germline.
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DOI:
10.1371/journal.pone.0020082
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发表时间:
2011
期刊:
影响因子:
3.7
通讯作者:
Ahringer J
Ahringer J
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Zeiser E;Frøkjær-Jensen C;Jorgensen E;Ahringer J

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在这里,我们描述了一个工具包,用于生产荧光标记蛋白在线虫种系和早期胚胎使用Mos1介导单拷贝插入(MosSCI)转化。我们已经将启动子和3‘端非编码区融合到不同的荧光蛋白序列上,产生了与MosSCI多点网关载体兼容的生殖系表达构建体。这些载体允许使用MosSCI将标记的转基因构建物作为单拷贝插入线虫基因组中的已知位置。我们还发现,两个线虫热休克启动子(Phsp-16.2和Phsp-16.41)可以通过MosSCI转化诱导转基因在胚系中的表达。这组灵活的新载体可供研究团体在质粒库中使用,应该会促进专注于线虫种系和早期胚胎的研究。
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.
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