RNA detection using peptide-inserted Renilla luciferase

被引：9

作者：

Andou, Takashi ^{[1
]}

Endoh, Tamaki ^{[2
]}

Mie, Masayasu ^{[1
]}

Kobatake, Eiry ^{[1
]}

机构：

[1] Tokyo Inst Technol, Dept Biol Informat, Grad Sch Biosci & Biotechnol, Midori Ku, Yokohama, Kanagawa 2268501, Japan

[2] Okayama Univ, Dept Biosci & Biotechnol, Grad Sch Nat Sci & Technol, Okayama 7008530, Japan

来源：

ANALYTICAL AND BIOANALYTICAL CHEMISTRY | 2009年 / 393卷 / 02期

关键词：

RNA detection; Complementation assay; Renilla luciferase; Arginine-rich motif peptide; RNA probe; PROTEIN-PROTEIN INTERACTIONS; LINKED MOLECULAR BEACONS; LIVING SUBJECTS; MESSENGER-RNA; CAENORHABDITIS-ELEGANS; MAJOR GROOVE; CELLS; COMPLEMENTATION; HYBRIDIZATION; COMPLEX;

D O I：

10.1007/s00216-008-2473-2

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

A novel complementation system with short peptide-inserted-Renilla luciferase (PI-Rluc) and split-RNA probes was constructed for noninvasive RNA detection. The RNA binding peptides HIV-1 Rev and BIV Tat were used as inserted peptides. They display induced fit conformational changes upon binding to specific RNAs and trigger complementation or discomplementation of Rluc. Split-RNA probes were designed to reform the peptide binding site upon hybridization with arbitrarily selected target RNA. This set of recombinant protein and split-RNA probes enabled a high degree of sensitivity in RNA detection. In this study, we show that the Rluc system is comparable to Fluc, but that its detection limit for arbitrarily selected RNA (at least 100 pM) exceeds that of Fluc by approximately two orders of magnitude.

引用

页码：661 / 668

页数：8

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