RNA detection using peptide-inserted Renilla luciferase
RNA detection using peptide-inserted Renilla luciferase
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DOI：
10.1007/s00216-008-2473-2
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发表时间：
2009-01-01
影响因子：
4.3
通讯作者：
Kobatake, Eiry
中科院分区：
文献类型：
--
作者：
Andou, Takashi;Endoh, Tamaki;Kobatake, Eiry
构建了一种新的基于短肽插入海肾荧光素酶（PI-Rluc）和分裂RNA探针的互补系统，用于非侵入性RNA检测。RNA结合肽HIV-1 Rev和BIV达特用作插入肽。它们在与特定RNA结合时显示诱导的适合构象变化，并触发Rluc的互补或去互补。分裂RNA探针被设计成在与任意选择的靶RNA杂交后改革肽结合位点。这组重组蛋白和分裂RNA探针能够在RNA检测中实现高度的灵敏度。在这项研究中，我们表明Rluc系统与Fluc相当，但其对任意选择的RNA（至少100 pM）的检测限超过Fluc约两个数量级。
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.