Analysis of intracellular localization of clock protein by means of bioluminescence imaging technique

生物发光成像技术分析时钟蛋白的细胞内定位

• 批准号: 21590266
• 负责人: NAKAJIMA Yoshihiro
• 金额: $ 3万
• 依托单位: National Institute of Advanced Industrial Science and Technology
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2011
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21590266/
• 关键词: 生物時計; ルシフェラーゼ; 発光イメージング; 核内移行

To visualize intracellular localization of clock protein by means of bioluminescence imaging technique, we have developed an enhanced green-emitting luciferase(ELuc) probe. ELuc exhibits a light signal in mammalian cells that is over 10-fold stronger than that of the firefly luciferase(FLuc), which is the most widely used luciferase reporter gene. We showed that ELuc produces a strong light signal in primary cells and tissues and that it enables the visualization of gene expression with high temporal resolution at the single-cell level. Moreover, we successfully imaged the nucleocytoplasmic shuttling of clock protein(mPER2 and mCRY2) by fusing ELuc at the intracellular level.

为了通过生物发光成像技术显示Clock蛋白在细胞内的定位，我们开发了一种增强型绿色发射荧光素酶(ELuc)探针。ELuc在哺乳动物细胞中显示出比萤火虫荧光素酶(Fluc)强10倍以上的光信号，后者是最广泛使用的荧光素酶报告基因。我们发现eLUC在原代细胞和组织中产生强烈的光信号，并且它能够在单细胞水平上以高时间分辨率可视化基因表达。此外，通过在细胞内水平融合ELuc，我们成功地成像了Clock蛋白(mPER2和mCRY2)的核质穿梭。

项目成果

Dual-Color Luciferase Mouse Directly Demonstrates Coupled Expression of Two Clock Genes

• DOI: 10.1021/bi100545h
• 发表时间: 2010-09-21
• 期刊: BIOCHEMISTRY
• 影响因子: 2.9
• 作者: Noguchi, Takako;Michihata, Tomoko;Nakajima, Yoshihiro
• 通讯作者: Nakajima, Yoshihiro

Enhanced red-emitting railroad worm luciferase for bioassays and bioimaging

• DOI: 10.1002/pro.279
• 发表时间: 2010-01-01
• 期刊: PROTEIN SCIENCE
• 影响因子: 8
• 作者: Li, Xueyan;Nakajima, Yoshihiro;Ohmiya, Yoshihiro
• 通讯作者: Ohmiya, Yoshihiro

Development of multicolor luciferase assay system using different coloremitting luciferases for monitoring multiple gene expressions

开发使用不同颜色发射荧光素酶的多色荧光素酶检测系统来监测多个基因表达

• 发表时间: 2009
• 作者: Oarada M.;et al.;下川哲昭;中島芳浩
• 通讯作者: 中島芳浩

Coactivation of the CLOCK-BMAL1 complex by CBP mediates resetting of the circadian clock

• DOI: 10.1242/jcs.070300
• 发表时间: 2010-10-15
• 期刊: JOURNAL OF CELL SCIENCE
• 影响因子: 4
• 作者: Lee, Yool;Lee, Jiwon;Kim, Kyungjin
• 通讯作者: Kim, Kyungjin

生物発光技術による細胞情報解析の新展開

利用生物发光技术进行细胞信息分析的新进展

• 发表时间: 2011
• 作者: 西谷正太;高村恒人;山下翔;藤澤隆史;篠原一之;中島芳浩
• 通讯作者: 中島芳浩