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Zoological Research
Kunming Institute of Zoology, Chinese Academy of Sciences
ISSN: 2095-8137
Vol. 29, No. 5, 2008, pp. 477-484
Bioline Code: zr08075
Full paper language: Chinese
Document type: Research Article
Document available free of charge

Zoological Research, Vol. 29, No. 5, 2008, pp. 477-484

 en Cloning, Expression and Sequence Analysis of A Luciferase Gene from the Chinese Firefly Pyrocoelia pygidialis check for this species in other resources
DONG, Ping-xuan; HOU, Qing-bai; LI, Xue-yan & LIANG, Xing-cai

Abstract

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 check for this species in other resources , Lampyris turkestanicus check for this species in other resources and Nyctophila cf. caucasica check for this species in other resources . 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.

Keywords
Pyrocoelia; Diurnal firefly; Pyrocoelia pygidialis; Luciferase; Homology modeling

 
© Copyright 2008 Kunming Institute of Zoology, the Chinese Academy of Sciences
Alternative site location: http://www.zoores.ac.cn/

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