基于phic31的果蝇荧光蛋白标记载体的模块化工具集。

IF 2.4 4区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Fly Pub Date : 2019-03-01 Epub Date: 2019-03-28 DOI:10.1080/19336934.2019.1595999
Jun Luo, Pingping Shen, Jiong Chen
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引用次数: 3

摘要

果蝇转基因技术和荧光蛋白融合是分析蛋白质表达模式、亚细胞定位和蛋白质动力学的有力工具。近年来,利用高效的philic31位点特异性整合系统对果蝇转基因技术进行了改进。许多新的和改进的荧光蛋白已被开发出来,具有令人满意的优点。然而,phi31系统和新开发的荧光蛋白尚未系统地应用于果蝇转基因载体。本研究基于phiC31位点特异性整合系统构建了c端荧光蛋白融合载体模块化工具集,用于果蝇转基因品系的生成。这些克隆载体包含多种荧光标签,包括蓝色、青色、绿色或红色荧光蛋白、光激活或光切换荧光蛋白、荧光计时器、光敏剂和双分子荧光互补标签。这些载体提供一系列转录调控选项,包括UAST、UASP、UASC、LexAop、QUAS、Ubi、αTub67C和αTub84B启动子,以及两种筛选标记选项,包括白色和朱红色基因。这些载体已经在体内进行了测试,可以产生功能性的荧光嵌合蛋白。
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A modular toolset of phiC31-based fluorescent protein tagging vectors for Drosophila.

The Drosophila transgenic technology and fluorescent protein fusions are powerful tools to analyze protein expression patterns, subcellular localization and protein dynamics. Recently, the Drosophila transgenic technology has been improved by the highly efficient phiC31 site-specific integration system. Many new and improved fluorescent proteins with desirable advantages have been developed. However, the phiC31 system and the newly developed fluorescent proteins have not been systematically applied in Drosophila transgenic vectors. Here, we have constructed a modular toolset of C-terminal fluorescent protein fusion vectors based on phiC31 site-specific integration system for the generation of transgenic Drosophila lines. These cloning vectors contain a variety of fluorescent tags, including blue, cyan, green or red fluorescent proteins, photoactivatable or photoswitchable fluorescent proteins, fluorescent timers, photosensitizers and bimolecular fluorescence complementation tags. These vectors provide a range of transcriptional regulation options including UAST, UASP, UASC, LexAop, QUAS, Ubi, αTub67C and αTub84B promoters, and two screening marker options including white and vermilion gene. The vectors have been tested in vivo and can produce fluorescent chimeric proteins that are functional.

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来源期刊
Fly
Fly 生物-生化与分子生物学
CiteScore
2.90
自引率
0.00%
发文量
17
审稿时长
>12 weeks
期刊介绍: Fly is the first international peer-reviewed journal to focus on Drosophila research. Fly covers a broad range of biological sub-disciplines, ranging from developmental biology and organogenesis to sensory neurobiology, circadian rhythm and learning and memory, to sex determination, evolutionary biology and speciation. We strive to become the “to go” resource for every researcher working with Drosophila by providing a forum where the specific interests of the Drosophila community can be discussed. With the advance of molecular technologies that enable researchers to manipulate genes and their functions in many other organisms, Fly is now also publishing papers that use other insect model systems used to investigate important biological questions. Fly offers a variety of papers, including Original Research Articles, Methods and Technical Advances, Brief Communications, Reviews and Meeting Reports. In addition, Fly also features two unconventional types of contributions, Counterpoints and Extra View articles. Counterpoints are opinion pieces that critically discuss controversial papers questioning current paradigms, whether justified or not. Extra View articles, which generally are solicited by Fly editors, provide authors of important forthcoming papers published elsewhere an opportunity to expand on their original findings and discuss the broader impact of their discovery. Extra View authors are strongly encouraged to complement their published observations with additional data not included in the original paper or acquired subsequently.
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