Molecular cloning and expression of the cDNAs encoding luciferin-regenerating enzyme from Luciola cruciata and Luciola lateralis.
In the firefly light organ, oxyluciferin, a product of the light-emitting reaction of firefly luciferase, is thought to be converted into luciferin. Previously, we isolated the luciferin-regenerating enzyme (LRE) from Photinus pyralis. LRE plays an important role in the recycling of oxyluciferin into luciferin. We have cloned two cDNAs encoding LRE, G-LRE and H-LRE, from poly(A)+ RNA of the lanterns of Luciola cruciata and Luciola lateralis, using reverse transcription-polymerase chain reaction, 5'-RACE (5'-rapid amplification of cDNA ends) and 3'-RACE. The putative translation products have molecular masses of 33,804 and 34,285 Da, corresponding to 309 and 307 amino acids, respectively. The deduced amino acid sequence of G-LRE shows 57 and 56% identity with H-LRE and A-LRE (P. pyralis), respectively. LRE (G-LRE, H-LRE, A-LRE) shows at most 39% amino acid sequence identity with insect anterior fat protein (AFP) and mammalian senescence marker protein-30 (SMP30). G-LRE and H-LRE were successfully expressed under the control of the lac promoter in Escherichia coli.
Gomi
Keiko
K
Research and Development Division, Kikkoman Corporation, 399 Noda, Noda-shi, 278-0037, Chiba, Japan. 8553@mail.kikkoman.co.jp
Hirokawa
Kozo
K
Kajiyama
Naoki
N
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D001517:Coleoptera / Q000201:enzymology / Q000235:genetics*; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D049454:Firefly Luciferin / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D007211:Indoles; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism; D008025:Ligases / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D011719:Pyrazines; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D007211:Indoles; D019476:Insect Proteins; D011719:Pyrazines; C014730:oxyluciferin; D049454:Firefly Luciferin; D008025:Ligases; C471161:luciferin-regenerating enzyme, insect