Mining the Sinorhizobium meliloti transportome to develop FRET biosensors for sugars, dicarboxylates and cyclic polyols.
Mining the Sinorhizobium meliloti transportome to develop FRET biosensors for sugars, dicarboxylates and cyclic polyols.
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DOI：
10.1371/journal.pone.0043578
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Poole PS
中科院分区：
文献类型：
--
作者：
Bourdès A;Rudder S;East AK;Poole PS
Förster共振能量转移（FRET）生物传感器是实时检测生物学重要的配体的强大工具，目前有fret双镜头可用于分布在八种化学物质中的22种化合物一个核苷酸酶，两个核苷酸，六个离子和三个植物雌激素）。 Meliloti Transportome Sinorhizobium tossytational筛选新的FRET生物传感器。
除编码FRET伴侣荧光蛋白之间的固定蛋白（SBP）的克隆基因开发了两个新的载体。和红色荧光蛋白（MKATE2）的fret伙伴。 D- Quilaticic酸，肌醇，L-rhamnose，l-糖果，β-二氨基葡萄糖（纤维二糖和刺激性），D-半乳糖糖和C4-二羧酸盐（苹果酸酯，琥珀酸酯，琥珀酸酯，草酸乙酸，我们所知），用于D- Quilaticic酸，L-rhamnose，L-rhamnose，Lhamnose，Lhamnose，Lhamnose，Lhamnose，Lhamnose，Lhamnose，Lhamnose，Lhamnose，Lhamnose，Lhamnose（Magobiose和gumarote）。描述基于三方ATP非依赖性周期（TRAP）传输系统的SBP的前两个FRET生物传感器构建体。
基于橙色（Morange2）和红色荧光蛋白（MKATE2）伙伴的FRET允许使用更长的波长光，从而使样品在较低的能量下更深入渗透，并随着本文所描述的货物增长而增加的分辨率。对于四种新的化合物；（i）环状多元醇，（ii）l-脱氧糖，（iii）β-连接的二糖和（iv）C4-二羧酸盐研究代谢在体内。
Förster resonance energy transfer (FRET) biosensors are powerful tools to detect biologically important ligands in real time. Currently FRET bisosensors are available for twenty-two compounds distributed in eight classes of chemicals (two pentoses, two hexoses, two disaccharides, four amino acids, one nucleobase, two nucleotides, six ions and three phytoestrogens). To expand the number of available FRET biosensors we used the induction profile of the Sinorhizobium meliloti transportome to systematically screen for new FRET biosensors. Two new vectors were developed for cloning genes for solute-binding proteins (SBPs) between those encoding FRET partner fluorescent proteins. In addition to a vector with the widely used cyan and yellow fluorescent protein FRET partners, we developed a vector using orange (mOrange2) and red fluorescent protein (mKate2) FRET partners. From the sixty-nine SBPs tested, seven gave a detectable FRET signal change on binding substrate, resulting in biosensors for D-quinic acid, myo-inositol, L-rhamnose, L-fucose, β-diglucosides (cellobiose and gentiobiose), D-galactose and C4-dicarboxylates (malate, succinate, oxaloacetate and fumarate). To our knowledge, we describe the first two FRET biosensor constructs based on SBPs from Tripartite ATP-independent periplasmic (TRAP) transport systems. FRET based on orange (mOrange2) and red fluorescent protein (mKate2) partners allows the use of longer wavelength light, enabling deeper penetration of samples at lower energy and increased resolution with reduced back-ground auto-fluorescence. The FRET biosensors described in this paper for four new classes of compounds; (i) cyclic polyols, (ii) L-deoxy sugars, (iii) β-linked disaccharides and (iv) C4-dicarboxylates could be developed to study metabolism in vivo.
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影响因子：
2.9
作者：
Bagaria, Ashima;Kumaran, Desigan;Burley, Stephen K.;Swaminathan, Subramanyam
通讯作者：
Swaminathan, Subramanyam
影响因子：
5.3
作者：
Fehr, M;Takanaga, H;Frommer, WB
通讯作者：
Frommer, WB
影响因子：
2.8
作者：
Galbraith, MP;Feng, SF;Rossbach, S
通讯作者：
Rossbach, S
DOI：
10.1073/pnas.142089199
发表时间：
2002-07-23
影响因子：
11.1
作者：
Fehr, M;Frommer, WB;Lalonde, S
通讯作者：
Lalonde, S
影响因子：
4.8
作者：
Fehr, M;Lalonde, S;Frommer, WB
通讯作者：
Frommer, WB