Magnetic resonance-based visualization of gene expression in mammalian cells using a bacterial polyphosphate kinase reporter gene
Magnetic resonance-based visualization of gene expression in mammalian cells using a bacterial polyphosphate kinase reporter gene
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DOI：
10.2144/000112319
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发表时间：
2007-02-01
期刊：
影响因子：
2.7
通讯作者：
Kokubo, Tetsuro
中科院分区：
文献类型：
--
作者：
Ki, Sewon;Sugihara, Fuminori;Kokubo, Tetsuro
基因表达报告系统，其中感兴趣的启动子克隆在一个易于检测的报告基因的上游，已经开发和广泛用于研究基因表达在原核生物和真核生物。不幸的是，大多数这些系统不能用于测定生物体内非表层组织中的基因表达。本研究研究了一个基于大肠杆菌多磷酸激酶（PPK）编码基因的新报告基因系统，该系统可用于监测哺乳动物细胞中的基因表达。PPK催化ATP合成无机多磷酸（polyP），由于哺乳动物细胞中不含有可检测水平的polyP，因此可以使用p -31磁共振波谱或p -31磁共振成像在哺乳动物细胞中测量PPK的活性。这里描述的ppk报告基因系统是非侵入性的，不需要外源底物，并且可以潜在地用于生物体的内部组织。
Gene expression reporter systems, in which a promoter of interest is cloned upstream of a readily assayed reporter gene, have been developed and used extensively to study gene expression in prokaryotes and eukaryotes. Unfortunately, most of these systems cannot be used to assay gene expression in nonsuperficial tissues in living organisms. This study examines a novel reporter gene system based on the gene encoding Escherichia coli polyphosphate kinase (PPK), which can be used to monitor gene expression in mammalian cells. PPK catalyzes the synthesis of inorganic polyphosphate (polyP) from ATP and because mammalian cells do not contain detectable levels of polyP PPK activity can be measured in mammalian cells using P-31-magnetic resonance spectroscopy or P-31-magnetic resonance imaging. The ppk reporter gene system described here is noninvasive, does not require an exogenous substrate, and can potentially be used in internal tissues of living organisms.