Cloning, Expression and Sequence Analysis of A Luciferase Gene from the Chinese Firefly Pyrocoelia pygidialis

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	Cloning, Expression and Sequence Analysis of A Luciferase Gene from the Chinese Firefly Pyrocoelia pygidialis
	Dong PX 1,2; Hou QB 1; Li XY 1; Liang XC[*]1
刊名	动物学研究
	2008
卷号	29 期号:5 页码:477-484
关键词	Pyrocoelia Diurnal firefly Pyrocoelia pygidialis Luciferase Homology modeling
其他题名	中国萤火虫云南窗萤荧光素酶cDNA的克隆表达和序列分析
通讯作者	liangxc@mail.kiz.ac.cn
合作状况	其它
中文摘要	从一种来自中国日行性萤火虫(云南窗萤)发光器官mRNA中克隆、测序并表达了有功能的荧光素酶.云南窗萤荧光素酶的cDNA序列有1647个碱基,编码548个氨基酸残基.从推测得到的氨基酸序列的比对分析得出:云南窗萤的荧光素酶与来自Lampyris noctiluca,L.turkestanicus和Nyctophila cf.caucasica三种萤火虫的荧光素酶有97.8%的序列一致性.从推测得出的氨基酸序列进行系统发育分析,其结果表明:云南窗萤和Lampyris+Nyctophila聚在一起,与同属的发光强夜行性的萤火虫不形成的单系.云南窗萤荧光素酶在大肠杆菌中表达的条带大约70kDa,并且在有荧光素存在时发出黄绿色荧光.对荧光素酶的结构模拟和分析表明,云南窗萤荧光素酶基因的氨基端和羧基端结构域之间的裂沟处存在这5个多肽环,这正是从其他荧光素酶推测得到的催化荧光反应时的底物结合位点.云南窗萤和窗萤属的其他3种萤火虫的荧光素酶卡目比,有13个不同氨基酸位点,位于模拟分子结构的表面.对于这些多肽环、不刚氨基酸残基和晶体结构的进一步研究有利于解释日行和夜行性萤火虫荧光素酶的差异.
英文摘要	The cDNA encoding the luciferase from lantern mRNA of one diurnal firefly Pyrocoelia pygidialis Pic, 1926 has been cloned, sequenced and functionally expressed. The cDNA sequence of P. pygidialis luciferase is 1647 base pairs in length, coding a protein of 548 amino acid residues. Sequence analysis of the deduced amino acid sequence showed that this luciferase had 97.8% resemblance to luciferases from the fireflies Lampyris noctiluca, Lampyris turkestanicus and Nyctophila cf. caucasica. Phylogenetic analysis using deduced amino acid sequence showed that P. pygidialis located at the base of Lampyris+Nyctophila clade with robust support (BP=97%); but did not show a monophyletic relationship with its congeneric species P. pectoralis, P. rufa and P. miyako, all three are strong luminous and nocturnal species. The expression worked in recombinant Escherichia coli. Expression product had a 70 kDa band and emitted yellow-green luminescence in the presence of luciferin. Five loops in the P. pygidialis luciferase, L1 (N198-G208), L2 (T240-G247), L3 (G317-K322), L4 (L343-I350) and L5 (G522-D532), were found from the structure modeling analysis in the cleft, where it was considered the active site for the substrate compound entering and binding. Different amino acid residues between the luciferases of P. pygidialis and the three other known strong luminous species can not explain the situation of weak or strong luminescence. Future study of these loops, residues or crystal structure analysis may be helpful in understanding the real differences between the luciferases between diurnal and nocturnal species.
收录类别	其他
资助信息	Supported partly by the Natural Foundation of Sciences of Yunnan Province (2006C0046Q); Partly by the Chinese Academy of Sciences (O706551141)
语种	英语
公开日期	2010-08-04
内容类型	期刊论文
源URL	[http://159.226.149.42:8088/handle/152453/915]
专题	昆明动物研究所_其他昆明动物研究所_基因起源组
作者单位	1.Kunming Institute of Zoology, the Chinese Academy of Sciences, Kunming Yunnan 650223, China 2.Graduate University of the Chinese Academy of Sciences, Beijing 100049, China
推荐引用方式 GB/T 7714	Dong PX,Hou QB,Li XY,et al. Cloning, Expression and Sequence Analysis of A Luciferase Gene from the Chinese Firefly Pyrocoelia pygidialis[J]. 动物学研究,2008,29(5):477-484.
APA	Dong PX,Hou QB,Li XY,&Liang XC[*].(2008).Cloning, Expression and Sequence Analysis of A Luciferase Gene from the Chinese Firefly Pyrocoelia pygidialis.动物学研究,29(5),477-484.
MLA	Dong PX,et al."Cloning, Expression and Sequence Analysis of A Luciferase Gene from the Chinese Firefly Pyrocoelia pygidialis".动物学研究 29.5(2008):477-484.