Expression of fluorescently tagged connexins: a novel approach to rescue function of oligomeric DsRed-tagged proteins

被引：76

作者：

Lauf, U ^{[1
]}

Lopez, P ^{[1
]}

Falk, MM ^{[1
]}

机构：

[1] Scripps Res Inst, Dept Cell Biol, La Jolla, CA 92037 USA

来源：

FEBS LETTERS | 2001年 / 498卷 / 01期

关键词：

green fluorescent protein; red fluorescent protein; gap junction; oligomeric membrane protein; aggregation of protein; protein chimera;

D O I：

10.1016/S0014-5793(01)02462-0

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：11 / 15

页数：5