Lampyris

Abstract: A full-length clone encoding Lampyris noctiluca (British glow-worm) luciferase was isolated from a complementary DNA (cDNA) expression library constructed with mRNA extracted from light organs. The luciferase was a 547-residue protein, as deduced from the nucleotide sequence. The protein was closely related to those of other lampyrid beetles, the similarity to Photinus pyralis luciferase being 84% and to Luciola 67%. In contrast, Lampyris luciferase had less sequence similarity to the luciferases of the click beetle Pyrophorus, at 48%. Engineering Lampyris luciferase in vitro showed that the C-terminal peptide containing 12 amino acids in Photinus and 9 amino acids in Lampyris was essential for bioluminescence. The pH optimum and the Km values for ATP and luciferin were similar for both Photinus and Lampyris luciferases, although the light emitted by the latter shifted towards the blue and was less stable at 37 °C. It was concluded that the molecular and biochemical properties were not sufficient to explain the glowing or flashing of the two beetles Lampyris and Photinus.

Abstract: Diaphanes is the fourth largest genus in Lampyridae, but no luciferase gene from this genus has been reported. In this paper, by PCR amplification of the genomic DNA, the luciferase gene of Diaphanes pectinealis, which is the first case from Diaphanes, was identified and sequenced. The luciferase gene from D. pectinealis spans 1958 base pairs (bp) from the start to the stop codon, including seven axons separated by six introns, and encoding a 547-residue-long polypeptide. Its deduced amino acid sequence showed high protein similarity to those of the Lampyrini tribe (93-94%) and the Cratomorphini tribe (92%), while low similarity was found with the North American firefly Photinus pyralis (83%) of the Photinini tribe within the same subfamily Lampyrinae. The phylogenetic analysis performed with the deduced amino acid sequences of the luciferase gene further confirms that D. pectinealis, Pyrocoelia, Lampyris, Cratomorphus, and Photinus belong to the same subfamily Lampyrinae, and Diaphanes is closely related to Pyrocoelia, Lampyris, and Cratomorphus. Furthemore, the phylogenetic analysis based on the nucleotide sequences of the luciferase gene indicates Diaphanes is a sister to Lampyris. The phylogenetic analyses are partly consistent with morphological (Branham & Wenzel, 2003) and mitochondrial DNA analyses (Li et al, 2006).

Abstract: The gene coding for beetle luciferase, the enzyme responsible for bioluminescence in over two thousand coleopteran species has, to date, only been characterized from one Palearctic species of Lampyridae. Here we report the characterization of the luciferase gene from a female beetle of an Iranian lampyrid species, Nyctophila cf. caucasica (Coleoptera:Lampyridae). The luciferase gene was composed of seven exons, coding for 547 amino acids, separated by six introns spanning 1976 bp of genomic DNA. The deduced amino acid sequences of the luciferase gene of N. caucasica showed 98.9% homology to that of the Palearctic species Lampyris noctiluca. Analysis of the 810 bp upstream region of the luciferase gene revealed three TATA boxes and several other consensus transcriptional factor recognition sequences presenting evidence for a putative core promoter region conserved in Lampyrinae from -190 through to -155 upstream of the luciferase start codon. Along with the core promoter region the luciferase gene was compared with orthologous sequences from other lampyrid species and found to have greatest identity to Lampyris turkistanicus and Lampyris noctiluca. The significant sequence identity to the former is discussed in relation to taxonomic issues of Iranian lampyrids.

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 978% 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