title: Molecular cloning and characterization of the nitric oxide synthase gene from kuruma shrimp, Marsupenaeus japonicus.
abstract: Nitric oxide (NO) signaling is involved in many physiological processes in vertebrates and invertebrates. In crustaceans, nitric oxide synthase (NOS) plays a significant role in the regulation of the nervous system and in innate immunity. Here, we describe the entire cDNA sequence (4616 bp) of the kuruma shrimp Marsupenaeus japonicus NOS (Mj NOS) generated using the reverse transcriptase-polymerase chain reaction (RT-PCR) and 5'- and 3'- rapid amplification PCRs of cDNA ends from brain and gill mRNAs. The open reading frame of Mj NOS encoded a protein of 1187 amino acids with an estimated mass of 134 kDa, and had an 82.3% sequence homology with the NOS gene of the land crab Gecarcinus lateralis. Highly conserved amino acid sequences in heme and tetrahydrobiopterin were observed in the oxygenase domain. FMN, FAD and NADPH were found in the reductase domain. Mj NOS mRNA was constitutively expressed in the brain, gill, intestine, thoracic ganglion and testis of the kuruma shrimp. When Vibrio penaeicida was injected into the kuruma shrimp, Mj NOS was expressed in the brain, gill, heart, lymphoid organ, intestine and thoracic ganglion. Mj NOS expression in the gill reached its peak 12 h and decreased to its normal level 24 h after V. penaeicida injection.
authors.affiliation: Graduate School of Agriculture, University of Miyazaki, 1-1, Gakuen Kibanadai-nishi, 889-2192 Miyazaki, Japan.
authors.forename: Mari
authors.initials: M
authors.lastname: Inada
authors.forename: Tohru
authors.initials: T
authors.lastname: Mekata
authors.forename: Raja
authors.initials: R
authors.lastname: Sudhakaran
authors.forename: Shogo
authors.initials: S
authors.lastname: Okugawa
authors.forename: Tomoya
authors.initials: T
authors.lastname: Kono
authors.forename: Amel Mohammed
authors.initials: AM
authors.lastname: El Asely
authors.forename: Nguyen T H
authors.initials: NT
authors.lastname: Linh
authors.forename: Terutoyo
authors.initials: T
authors.lastname: Yoshida
authors.forename: Masahiro
authors.initials: M
authors.lastname: Sakai
authors.forename: Toshifumi
authors.initials: T
authors.lastname: Yui
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Itami
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D020869:Gene Expression Profiling; D015971:Gene Expression Regulation, Enzymologic; D007113:Immunity, Innate; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019001:Nitric Oxide Synthase / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D033561:Penaeidae / Q000145:classification / Q000201:enzymology* / Q000276:immunology / Q000378:metabolism / Q000382:microbiology; D010802:Phylogeny; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D014733:Vibrio / Q000502:physiology
chemical_list: D019001:Nitric Oxide Synthase