Expression of fluorescently tagged connexins: a novel approach to rescue function of oligomeric DsRed-tagged proteins
Expression of fluorescently tagged connexins: a novel approach to rescue function of oligomeric DsRed-tagged proteins
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DOI：
10.1016/s0014-5793(01)02462-0
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发表时间：
2001-06-01
期刊：
影响因子：
3.5
通讯作者：
Falk, MM
中科院分区：
文献类型：
--
作者：
Lauf, U;Lopez, P;Falk, MM
DsRed 是一种新型的亮红色荧光蛋白，最近上市，为与绿色荧光蛋白 (GFP) 结合的双标记和荧光共振电子转移实验提供了广泛的实验机会。与 GFP 不同的是，用 DsRed 标记的蛋白质经常被发现在细胞内聚集。在这里，我们报告了一种简单的方法，可以挽救带有 DsRed 标记的寡聚蛋白的功能，我们证明了这种方法在寡聚膜通道的亚基蛋白、间隙连接蛋白上的可行性。此外，转染后 12-16 小时很容易检测到 DsRed 荧光，比以前报道的要早得多，并且可以很容易地与共表达的 GFP 区分开来，因此，这种方法可以消除这种高度有吸引力的自发荧光蛋白的主要缺点。 (C) 2001 年欧洲生化学会联合会。由 Elsevier Science B.V. 出版。保留所有权利。
A novel, brilliantly red fluorescent protein, DsRed has become available recently opening up a wide variety of experimental opportunities for double labeling and fluorescence resonance electron transfer experiments in combination with green fluorescent protein (GFP). Unlike in the case of GFP, proteins tagged,vith DsRed were often found to aggregate within the cell. Here we report a simple method that allows rescuing the function of an oligomeric protein tagged with DsRed, We demonstrate the feasibility of this approach on the subunit proteins of an oligomeric membrane channel, gap junction connexins, Additionally, DsRed fluorescence was easily detected 12-16 h post transfection, much earlier than previously reported, and could readily be differentiated from co-expressed GFP, Thus, this approach can eliminate the major drawbacks of this highly attractive autofluorescent protein. (C) 2001 Federation of European Biochemical Societies. Published by Elsevier Science B.V. All rights reserved.