A modular toolset for recombination transgenesis and neurogenetic analysis of Drosophila.
A modular toolset for recombination transgenesis and neurogenetic analysis of Drosophila.
复制标题
DOI：
10.1371/journal.pone.0042102
复制
发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
McCabe BD
中科院分区：
文献类型：
--
作者：
Wang JW;Beck ES;McCabe BD
转基因果蝇除了是人类神经系统疾病的有价值的模型之外，还为我们理解神经系统发育、生理和行为做出了广泛的贡献。在这里，我们生成了一系列新型模块化转基因载体，旨在优化和加速果蝇转基因的生产和分析。我们构建了一种新的载体骨架pBID，其允许phiC31靶向转基因整合并掺入绝缘子序列以确保特异性和均匀的转基因表达。在此框架下，我们已经构建了一系列构建体，这些构建体或者与现有的基于限制性酶的载体向后兼容，或者利用Gateway重组技术进行高通量克隆。这些载体允许内源启动子或Gal 4靶向表达具有或不具有荧光蛋白或表位标签的转基因蛋白。此外，我们已经产生了促进转基因剪接异构体特异性RNA抑制基因表达的构建体。我们证明了这些结构的实用性，以分析参与神经系统发育，生理和神经退行性疾病的蛋白质。我们期望这些试剂将促进果蝇遗传分析在神经系统和其他组织中的熟练度和复杂性。
Transgenic Drosophila have contributed extensively to our understanding of nervous system development, physiology and behavior in addition to being valuable models of human neurological disease. Here, we have generated a novel series of modular transgenic vectors designed to optimize and accelerate the production and analysis of transgenes in Drosophila. We constructed a novel vector backbone, pBID, that allows both phiC31 targeted transgene integration and incorporates insulator sequences to ensure specific and uniform transgene expression. Upon this framework, we have built a series of constructs that are either backwards compatible with existing restriction enzyme based vectors or utilize Gateway recombination technology for high-throughput cloning. These vectors allow for endogenous promoter or Gal4 targeted expression of transgenic proteins with or without fluorescent protein or epitope tags. In addition, we have generated constructs that facilitate transgenic splice isoform specific RNA inhibition of gene expression. We demonstrate the utility of these constructs to analyze proteins involved in nervous system development, physiology and neurodegenerative disease. We expect that these reagents will facilitate the proficiency and sophistication of Drosophila genetic analysis in both the nervous system and other tissues.
登录
查看更多内容
影响因子：
56.9
作者：
Kwiatkowski, T. J., Jr.;Bosco, D. A.;Brown, R. H., Jr.
通讯作者：
Brown, R. H., Jr.
影响因子：
64.8
作者：
Graveley BR;Brooks AN;Carlson JW;Duff MO;Landolin JM;Yang L;Artieri CG;van Baren MJ;Boley N;Booth BW;Brown JB;Cherbas L;Davis CA;Dobin A;Li R;Lin W;Malone JH;Mattiuzzo NR;Miller D;Sturgill D;Tuch BB;Zaleski C;Zhang D;Blanchette M;Dudoit S;Eads B;Green RE;Hammonds A;Jiang L;Kapranov P;Langton L;Perrimon N;Sandler JE;Wan KH;Willingham A;Zhang Y;Zou Y;Andrews J;Bickel PJ;Brenner SE;Brent MR;Cherbas P;Gingeras TR;Hoskins RA;Kaufman TC;Oliver B;Celniker SE
通讯作者：
Celniker SE
DOI：
10.1523/jneurosci.3717-11.2012
发表时间：
2012-05-16
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Beck ES;Gasque G;Imlach WL;Jiao W;Jiwon Choi B;Wu PS;Kraushar ML;McCabe BD
通讯作者：
McCabe BD
影响因子：
14.9
作者：
KLEMENZ, R;WEBER, U;GEHRING, WJ
通讯作者：
GEHRING, WJ
影响因子：
34.7
作者：
Bellen, Hugo J.;Tong, Chao;Tsuda, Hiroshi
通讯作者：
Tsuda, Hiroshi