Molecular probes to visualize cellular processes in vivo

分子探针使体内细胞过程可视化

• 批准号: 23655152
• 负责人: SATO Moritoshi
• 金额: $ 2.58万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23655152/
• 关键词: 生体機能関連化学; 生物発光; 細胞; 生体; イメージング; プローブ; ルシフェラーゼ; ガン細胞; 転移; 可視化; エネルギー移動; 蛍光

In this study, a rational development of superluminescent variants of a luciferase derived from Gaussia princeps was demonstrated. A putative catalytic center of Gaussia luciferase was assigned and modified by a site-directed mutagenesis. The potent variants were found to generate up to 10 times stronger bioluminescence, emitting red shifts of up to 33 nm with natural coelenterazine than native GLuc, rendering an efficient optical signature in bioassays. The advantageous properties were demonstrated with metastases of murine B16 melanoma in BALB/c nude mice.

在这项研究中，合理地开发了一种来自赤霉菌的荧光素酶的超发光变体。通过定点突变，确定了赤子荧光素酶的一个可能的催化中心。研究发现，这些有效的变异体可以产生高达10倍的生物发光，与天然的GLuc相比，天然的Coelentazine发出的红移高达33 nm，在生物检测中呈现出高效的光学特征。以BALB/c裸鼠BALB/c黑色素瘤的转移为例，证实了该方法的优越性。