Center for Ecological and Environmental Sciences, Key Laboratory for Space Bioscience & Biotechnology, Northwestern Polytechnical University, Xi'an, China.
State Key Laboratory of Genetic Resources and Evolution, Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, China.
Photochem Photobiol. 2019 Sep;95(5):1186-1194. doi: 10.1111/php.13109. Epub 2019 May 9.
Lamprigera (Lampyridae) is a small genus with only 17 species distributing in Asian countries. Its larviform females and alate males can produce continuously strong yellow-green light at night. However, no luciferase gene was reported for this genus and its subfamily-level phylogenetic position still remains uncertain. Here, we cloned the luciferase gene from one Chinese species, Lamprigera yunnana, by reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). This luciferase includes 551 deduced amino acids (AA) with the sequence identity of 71.8-76.8%, 67.5-70.7%, 68.4-75.3%, 77.8% and 59.5% to those from Lampyrinae, Luciolinae, Ototretinae, Cyphonocerinae and Photurinae, respectively. Phylogenetic analyses of deduced AA of luciferases suggest that Lamprigera locates outside Lampyrinae, in which it was originally placed in traditional taxonomy. The luciferase was produced in vitro as recombinant protein, and its biochemical properties were characterized. It possesses significant luminescence activity at pH 7.8, and its K for D-luciferin and ATP are 61 μm and 122 μm, respectively. It shows the highest activity at 37°C and is completely inactivated at 55°C. It is pH-sensitive with the maximum emission spectrum of 566 nm at pH 7.8. Our data provide new insights into Lamprigera luciferase and its phylogenetic position.
灯萤科（Lampyridae）是一个小属，仅有 17 种分布于亚洲国家。其幼虫状的雌性和有翅的雄性在夜间能持续产生强烈的黄绿色光。然而，该属及其亚科的系统发育位置仍不确定，尚未报道其荧光素酶基因。本研究通过反转录-聚合酶链式反应（RT-PCR）和快速扩增 cDNA 末端（RACE），从中国的一种灯萤（Lamprigera yunnana）克隆出荧光素酶基因。该荧光素酶包含 551 个推导的氨基酸（AA），与 Lampyrinae、 Luciolinae、Ototretinae、Cyphonocerinae 和 Photurinae 的序列同一性分别为 71.8-76.8%、67.5-70.7%、68.4-75.3%、77.8%和 59.5%。推导的 AA 系统发育分析表明，灯萤科位于灯萤科之外，在传统分类学中，它最初被放置在该科中。该荧光素酶在体外作为重组蛋白产生，并对其生化特性进行了表征。它在 pH 7.8 时具有显著的发光活性，其对 D-荧光素和 ATP 的 K 值分别为 61μm 和 122μm。它在 37°C 时表现出最高的活性，在 55°C 时完全失活。它对 pH 敏感，最大发射光谱在 pH 7.8 时为 566nm。我们的数据为灯萤科荧光素酶及其系统发育地位提供了新的见解。
