Gene Cloning and Functional Analysis of the Luciferase from Luminous Syllids of the Genus Odontosyllis
Gene Cloning and Functional Analysis of the Luciferase from Luminous Syllids of the Genus Odontosyllis
复制标题
齿鳅属发光鳅荧光素酶基因克隆及功能分析
DOI：
10.1007/978-1-0716-2453-1_1
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Ohmiya Yoshihiro
中科院分区：
文献类型：
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作者：
Yasuno Rie;Mitani Yasuo;Ohmiya Yoshihiro
海洋萤火虫齿形虫。产生蓝绿色生物发光(BL)。尽管多年来的研究，这种独特的荧光素-荧光素酶反应的分子机制还没有被阐明。最近，编码十一齿假单胞菌和红假单胞菌荧光素酶的基因已被鉴定出来。在这里，我们描述了蜻蜓荧光素酶的基因克隆技术。从少量样品中进行高灵敏的质谱分析并结合RNA测序。利用在哺乳动物细胞中表达的重组蛋白，在体外用BL法证实了克隆的cDNA的荧光素酶活性。
The marine firewormOdontosyllisspp. produce the bluish-green bioluminescence (BL). Despite years of research, molecular mechanisms of this unique luciferin–luciferase reaction have not been elucidated. Recently, the genes encoding luciferases ofO.undecimdontaandO.enoplahave been identified. Here, we describe gene cloning techniques for the luciferase ofOdontosyllisspp. from a small number of specimens using highly sensitive mass spectrometry analysis in combination with RNA-sequencing. The luciferase activities of the cloned cDNAs are confirmed by BL assay in vitro using recombinant protein expressed in mammalian cells.