a modular toolset for recombination transgenesis and neurogenetic analysis of drosophila模块化重组转基因技术工具集和neurogenetic果蝇的分析.pdfVIP

A Modular Toolset for Recombination Transgenesis and Neurogenetic Analysis of Drosophila Ji-Wu Wang, Erin S. Beck, Brian D. McCabe* Center for Motor Neuron Biology and Disease, Department of Pathology and Cell Biology, Department of Neuroscience, Columbia University, New York, New York, United States of America Abstract 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. Citation: Wang J-W, Beck ES, McCabe BD (2012) A Modular Toolset for Recombination Transgenesis and Neurogenetic Analysis of Drosophila. PLoS ONE 7(7): e42102. doi:10.1371/journal.p