Molecular cloning and characterization of the nitric oxide synthase gene from kuruma shrimp, Marsupenaeus japonicus.
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.
Inada
Mari
M
Graduate School of Agriculture, University of Miyazaki, 1-1, Gakuen Kibanadai-nishi, 889-2192 Miyazaki, Japan.
Mekata
Tohru
T
Sudhakaran
Raja
R
Okugawa
Shogo
S
Kono
Tomoya
T
El Asely
Amel Mohammed
AM
Linh
Nguyen T H
NT
Yoshida
Terutoyo
T
Sakai
Masahiro
M
Yui
Toshifumi
T
Itami
Toshiaki
T
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
D019001:Nitric Oxide Synthase