Reassembling green fluorescent protein for in vitro evaluation of trans-translation
Reassembling green fluorescent protein for in vitro evaluation of trans-translation
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DOI：
10.1093/nar/gkz1204
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发表时间：
2020-02-28
影响因子：
14.9
通讯作者：
Gillet, Reynald
中科院分区：
文献类型：
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作者：
Guyomar, Charlotte;Thepaut, Marion;Gillet, Reynald
为了发现具有更高活性和选择性的新抗生素，我们建立了一个可靠的体外报告系统来检测反翻译活性，这是细菌中停止在问题信使RNA （mRNA）上的核糖体循环的主要机制。该系统是基于一种工程tmRNA变体，该变体在反翻译活跃时重组绿色荧光蛋白（GFP）。我们的系统适用于荧光高通量筛选化合物。
In order to discover new antibiotics with improved activity and selectivity, we created a reliable in vitro reporter system to detect trans-translation activity, the main mechanism for recycling ribosomes stalled on problematic messenger RNA (mRNA) in bacteria. This system is based on an engineered tmRNA variant that reassembles the green fluorescent protein (GFP) when trans-translation is active. Our system is adapted for high-throughput screening of chemical compounds by fluorescence.