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巴西独特隐翅萤幼虫萤火虫荧光素酶cDNA的克隆与特性分析:与欧洲黄缘萤和亚洲胸窗萤荧光素酶的相似性

Cloning and characterization of the cDNA for the Brazilian Cratomorphus distinctus larval firefly luciferase: similarities with European Lampyris noctiluca and Asiatic Pyrocoelia luciferases.

作者信息

Viviani V R, Arnoldi Frederico G C, Brochetto-Braga M, Ohmiya Y

机构信息

Departamento de Biologia, Instituto de Biociências, Universidade Estadual de São Paulo, Av. 24A, 1515, Bela Vista, Rio Claro, SP 13506-900, Brazil.

出版信息

Comp Biochem Physiol B Biochem Mol Biol. 2004 Oct;139(2):151-6. doi: 10.1016/j.cbpc.2004.05.012.

Abstract

Studies on firefly (Lampyridae) luciferases have focused on nearctic species of Photinus and Photuris and Euroasiatic species of Lampyris, Luciola, Hotaria, and Pyrocoelia. Despite accounting for the greatest diversity of fireflies in the world, no molecular studies have been carried out on the highly diverse genera from the neotropical region. Here we report the luciferase cDNA cloning for the larva of the Brazilian firefly Cratomorphus distinctus. The cDNA has 1978 bp and codes for a 547-residue-long polypeptide. Noteworthy, sequence comparison as well as functional properties show the highest degree of similarity with Lampyris noctiluca (93%) and Pyrocoelia spp. (91%) luciferases, suggesting a close phylogenetic relationship despite the geographical distance separating these species. The bioluminescence emission spectrum peaks at 550 nm and, as expected, is sensitive to pH, shifting to 605 nm at pH 6. The kinetic properties of the recombinant luciferase were similar to those of other firefly luciferases.

摘要

对萤火虫(萤科)荧光素酶的研究主要集中在北美洲的Photinus和Photuris属以及欧亚大陆的Lampyris、Luciola、Hotaria和Pyrocoelia属。尽管新热带地区的萤火虫种类最为丰富,但尚未对该地区高度多样化的属进行分子研究。在此,我们报告了巴西萤火虫Cratomorphus distinctus幼虫的荧光素酶cDNA克隆。该cDNA有1978 bp,编码一个由547个氨基酸组成的多肽。值得注意的是,序列比较和功能特性显示,它与夜光萤(93%)和Pyrocoelia属荧光素酶(91%)具有最高程度的相似性,这表明尽管这些物种在地理上相隔甚远,但它们在系统发育上关系密切。生物发光发射光谱在550 nm处达到峰值,正如预期的那样,它对pH敏感,在pH 6时会移至605 nm。重组荧光素酶的动力学特性与其他萤火虫荧光素酶相似。

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