Highlighted generation of fluorescence signals using simultaneous two-color irradiation on Dronpa mutants
Highlighted generation of fluorescence signals using simultaneous two-color irradiation on Dronpa mutants
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DOI：
10.1529/biophysj.107.105882
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发表时间：
2007-06-01
影响因子：
3.4
通讯作者：
Miyawaki, Atsushi
中科院分区：
文献类型：
--
作者：
Ando, Ryoko;Flors, Cristina;Miyawaki, Atsushi
Dronpa吸收蓝光并发出明亮的绿色荧光。它也可以通过490 nm的强辐射转换为非荧光状态，然后可以通过400 nm的辐射切换回原始发射状态。通过半随机诱变研究，我们已经开发出两个突变体的Dronpa，显示有效的光开关动力学。与Dronpa相比，突变体可以更有效地被蓝光关闭。因此，用氩激光线（488 nm）激发使突变体迅速变暗，以致不能观察到实质性的荧光信号。用紫色激光二极管（405 nm）激发也不产生荧光信号。然而，同时488-和405-nm照射导致两种状态之间的快速振荡，从而保持发射状态的人口足够大，以产生足够明亮的荧光信号。
Dronpa absorbs blue light and emits bright green fluorescence. It can also be converted by strong irradiation at 490 nm to a non fluorescent state, which can then be switched back to the original emissive state with irradiation at 400 nm. Through semirandom mutagenesis studies, we have developed two mutants of Dronpa that show efficient photoswitching kinetics. Compared to Dronpa, the mutants can be turned off by blue light more efficiently. Thus, excitation with an argon laser line (488 nm) makes the mutants quickly become dark such that no substantial fluorescence signals can be observed. Excitation with a violet laser diode (405 nm) also produces no fluorescence signals. Simultaneous 488- and 405-nm irradiation, however, results in a rapid oscillation between the two states, thereby keeping the emissive state population large enough to produce sufficiently bright fluorescence signals.