Site-specific biosynthetic incorporation of a fluorescent tag into proteins via Cysteine-tRNACys

被引：4
|
作者：
Lien, L [1 ]
Ananda, P [1 ]
Seneviratne, K [1 ]
Jaikaran, ASI [1 ]
Woolley, GA [1 ]
机构：
[1] Univ Toronto, Dept Chem, Toronto, ON M5S 3H6, Canada
来源：
ANALYTICAL BIOCHEMISTRY | 2002年 / 307卷 / 02期
基金：
加拿大健康研究院;
关键词：
fluorescence; tRNA; cysteine; probe; protein; labeling; modification;
D O I：
10.1016/S0003-2697(02)00035-0
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Site-specific incorporation of biophysical probes into proteins during translation can permit structure/function studies on selected proteins in heterogeneous environments. We report here a procedure for incorporating a fluorescent tag into proteins via Escherichia coli Cys-tRNA(Cys) during in vitro protein synthesis. Naturally occurring Cys-tRNA(Cys) is an attractive vehicle for fluorophore incorporation since it can be readily prepared in quantity and reacted with commercially available fluorophores. Moreover, proteins can often be constructed with a single Cys so that fluorophore incorporation results in a tag at a unique site. (C) 2002 Elsevier Science (USA). All rights reserved.
引用
收藏
页码：252 / 257
页数：6
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