摘要
目的建立一种基于Diels-Alder环加成反应用于活细胞内量子点(quantum dots,QDs)标记蛋白质的新方法。方法反式环辛烯(trans-cyclooctene 2,TCO2)修饰的类转录激活因子效应物(transcription activator-like effectors,TALE)蛋白与偶联四嗪(tetrazine 1,Tz1)的红色量子点QD625在Vero活细胞内特异性结合,形成量子点荧光标记的蛋白TALE-QD625,通过激光共聚焦显微镜观察TALE-QD625在Vero细胞内的荧光信号分布。结果基于Tz1和TCO2的Diels-Alder环加成反应在Vero活细胞内将QD625标记至TALE蛋白,并随TALE进入细胞核内,在核内观察到红色的QDs荧光信号,成功建立活细胞内量子点标记蛋白方法。结论通过Diels-Alder环加成反应首次实现活细胞内量子点标记蛋白,为蛋白单分子成像提供一种新的标记方法。
Objective To establish a new method of labeling a protein with quantum dots( QDs) within live cells via the Diels-Alder cycloaddition. Methods Transcription activator-like effectors( TALE) decorated with trans-cyclooctene TCO2 and tetrazine 1( Tz1)-conjugated QD625 were combined specially in the live Vero cells to yield a QD625-labelled TALE( TALE-QD625). The distribution of fluorescent signal for TALE-QD625 in Vero cells were observed through confocal microscopy. Results The QD625 was conjugated with TALE based on the Tz1 and TCO2 Diels-Alder cycloaddition labeling system and subsequently entered the cell nucleus. Red fluorescent signal of TALE-QD625 was observed in the nucleus of Vero cells. Conclusion It is the first time that QDs have been used to specially label a protein via the Diels-Alder cycloaddition within live cells,which will provide a new labeling method for single protein molecule imaging.
作者
王明秀
张加玲
刘云凤
乔增杰
WANG Mingxiu ZHANG Jialing LIU Yunfeng QIAO Zengjie(Department of Sanitary Inspection, School of Public Health, Shanxi Medical University, Taiyuan 030001, China)
出处
《山西医科大学学报》
CAS
2017年第9期895-899,共5页
Journal of Shanxi Medical University
基金
病毒学国家重点实验室开放研究基金资助项目(2017IOV005)
山西医科大学青年基金资助项目(02201512)