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 bas...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 tufa and P miyako, all three are strong luminous and nocturnal species. The expression worked in recombinant Escherichia coli. Expression product had a 70kDa band and emitted yellow-green luminescence in the presence of luciferin. Five loops in the P pygidialis luciferase, L1 （NI98-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.展开更多
Morphology of different development stages from egg, larva, pupa to adult （male and female） of the firefly, Pyrocoelia pygidialis Pic, was described in details. Some biological habits were noted according to the rec...Morphology of different development stages from egg, larva, pupa to adult （male and female） of the firefly, Pyrocoelia pygidialis Pic, was described in details. Some biological habits were noted according to the records of field collection and observation, with references to the rearing in the laboratory. With weakly luminous egg, larva, pupa and female adult, and much weaker luminous male adult only when disturbed, P pygidialis Pic is considered as a diurnal firefly, and chemical signal, instead of light signal, is possibly its dominant means of male-female recognition.展开更多
基金the Natural Foundation of Sciences of Yunnan Province (2006C0046Q)Partly by the Chinese Academy of Sciences (O706551141)
文摘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 tufa and P miyako, all three are strong luminous and nocturnal species. The expression worked in recombinant Escherichia coli. Expression product had a 70kDa band and emitted yellow-green luminescence in the presence of luciferin. Five loops in the P pygidialis luciferase, L1 （NI98-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.
基金This project was supported by the Natural Scientific Foundation of Yunnan Province (2006C0046Q).
文摘Morphology of different development stages from egg, larva, pupa to adult （male and female） of the firefly, Pyrocoelia pygidialis Pic, was described in details. Some biological habits were noted according to the records of field collection and observation, with references to the rearing in the laboratory. With weakly luminous egg, larva, pupa and female adult, and much weaker luminous male adult only when disturbed, P pygidialis Pic is considered as a diurnal firefly, and chemical signal, instead of light signal, is possibly its dominant means of male-female recognition.
