Lauf U, Lopez P, Falk M M
Department of Cell Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
FEBS Lett. 2001 Jun 1;498(1):11-5. doi: 10.1016/s0014-5793(01)02462-0.
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 with 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.
一种新型的、色泽鲜艳的红色荧光蛋白DsRed最近问世，为与绿色荧光蛋白（GFP）结合进行双标记和荧光共振电子转移实验开辟了广泛的实验机会。与GFP的情况不同，用DsRed标记的蛋白质常常在细胞内聚集。在此，我们报告一种简单的方法，可挽救用DsRed标记的寡聚蛋白的功能。我们在寡聚膜通道——间隙连接连接蛋白的亚基蛋白上证明了这种方法的可行性。此外，转染后12 - 16小时就能轻松检测到DsRed荧光，比之前报道的要早得多，并且能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极具吸引力的自发荧光蛋白的主要缺点。
