One- and two-photon photoactivation of a paGFP-fusion protein in live Drosophila embryos
One- and two-photon photoactivation of a paGFP-fusion protein in live Drosophila embryos
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DOI：
10.1016/j.febslet.2004.11.092
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发表时间：
2005-01-17
期刊：
影响因子：
3.5
通讯作者：
Arndt-Jovin, DJ
中科院分区：
文献类型：
--
作者：
Post, JN;Lidke, KA;Arndt-Jovin, DJ
我们构建了一个可光激活的果蝇组蛋白2A变体绿色荧光融合蛋白（H2 AvDpaGFP），用于跟踪果蝇胚胎中的染色质位点.通过在408 nm处的单光子二极管激光照射实现了paGFP的激活，但是激活的核未能分裂。光转换也可以通过在780-840 nm范围内的双光子fs脉冲来实现。整装胚胎的活力只能在820 nm处维持，在该波长处，我们可以激活、同时跟踪和定量多个荧光位点的迁移率。本报告构成了双光子激活的paGFP和使用的paGFP融合蛋白在整个生物体的调查的第一个演示。(C)2004年欧洲生物化学学会联合会。Elsevier B. V.出版，保留所有权利。
We constructed a photoactivatable Drosophila histone 2 A variant green fluorescent fusion protein (H2AvDpaGFP) for tracking chromatin loci in living Drosophila embryos. Activation of paGFP was achieved by irradiation from a single-photon diode laser at 408 nm, but activated nuclei failed to divide. Photoconversion could also be achieved by two-photon fs pulses in the range of 780-840 nm. Viability in whole-mount embryos could only be maintained at 820 nm, at which we could activate, simultaneously track and quantitate the mobility of multiple fluorescent loci. This report constitutes the first demonstration of two-photon activation of paGFP and the use of a paGFP-fusion protein in investigations of whole organisms. (C) 2004 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.