期刊论文

A Fluorescence Enhancement Method for Real-Time Observation of Ca~(2+) Signaling Transduction in BY2 Cells

中文

中国细胞生物学学报

1674-7666

2018

40

11

1858-1866

10.11844/cjcb.2018.11.0147

湖南省研究生科研创新项目（批准号:CX2016B296）资助的课题~~；

本校为第一机构

钙离子作为第二信使是细胞信号转导的重要成员,也是细胞信号转导研究的重要对象.为了更有效地实时观察植物细胞中的钙离子信号,将响应钙离子的水母素(aequorin,AEQ)基因转化到已由绿色荧光蛋白(GFP)标记的分泌载体膜蛋白SCMP2(secretive carrier membrane proteins 2)的烟草BY2细胞(SCAMP2-GFP).经过对转化细胞的筛选和分子检测,获得了双价转化的BY2细胞株.转化细胞经腔肠素处理并实时显微观察,能较清晰地观察到细胞中钙离子响应的荧光信号,水母素单价转化细胞则几乎观察不到荧光信号,说明水母素响应钙离子并作用于腔肠素所发弱荧光能进一步激发GFP荧光,使弱荧光有效增强.尽管响应钙离子的双价自...

Calcium ion as the second messenger is an important member of cell signal transduction and is also a focused object of cell signal transduction research. It is meaningful to develop a method more sensitive and dynamical for real-time observation of calcium ion signal in plant cells to study the cellular signal transduction. In this paper, the gene of a calcium ion responsible aequorin(AEQ) original from jellyfish was transformed into tobacco BY2(bright yellow 2) cell that was SCMP2(secretive carrier membrane proteins 2) labeled with green fluorescent protein(SCAMP2-GFP). The bivalent transgeni...