[红外荧光蛋白iRFP作为荧光共振能量转移的受体]

[Infrared Fluorescent Protein iRFP as an Acceptor for Förster Resonance Energy Transfer].

作者信息

Zlobovskaya O A, Sarkisyan K S, Lukyanov K A

出版信息

Bioorg Khim. 2015 May-Jun;41(3):299-304. doi: 10.1134/s1068162015030139.

DOI:10.1134/s1068162015030139

Abstract

Bacteriophytochrome-based infrared fluorescent protein iRFP was tested as an acceptor for F6rster resonance energy transfer (FRET). Far-red GFP-like fluorescent proteins mKate2, eqFP650, and eqFP670 were used as donors; Bacterial expression vectors encoding donor and acceptor proteins fused by a 17-amino acid linker were.constructed. FRET for purified proteins in vitro was, estimated from increase of the donor emission after digestion of the linker. Among the three constructs tested, the most efficient FRET (approximately 30%) was detected for the eqFP650-iRFP pair.

摘要

基于细菌光敏色素的红外荧光蛋白iRFP被测试作为Förster共振能量转移（FRET）的受体。远红绿色荧光蛋白类似物mKate2、eqFP650和eqFP670被用作供体；构建了编码通过17个氨基酸接头融合的供体和受体蛋白的细菌表达载体。体外纯化蛋白的FRET是根据接头消化后供体发射的增加来估计的。在测试的三种构建体中，eqFP650-iRFP对检测到最有效的FRET（约30%）。