Graduate School of Agriculture, University of Miyazaki, 1-1, Gakuen Kibanadai-nishi, 889-2192 Miyazaki, Japan.
Fish Shellfish Immunol. 2010 Apr;28(4):701-11. doi: 10.1016/j.fsi.2010.01.010. Epub 2010 Jan 28.
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.
一氧化氮（NO）信号参与脊椎动物和无脊椎动物的许多生理过程。在甲壳动物中，一氧化氮合酶（NOS）在神经系统和先天免疫的调节中起着重要作用。在这里，我们使用逆转录聚合酶链反应（RT-PCR）和脑和鳃 mRNA 的 5'和 3'快速扩增 cDNA 末端从脑和鳃 mRNA 中生成了斑节对虾 Marsupenaeus japonicus NOS（Mj NOS）的全长 cDNA 序列（4616 bp）。Mj NOS 的开放阅读框编码了一种 1187 个氨基酸的蛋白质，估计分子量为 134 kDa，与陆地蟹 Gecarcinus lateralis 的 NOS 基因具有 82.3%的序列同源性。在加氧酶结构域中观察到血红素和四氢生物蝶呤的高度保守氨基酸序列。在还原酶结构域中发现了 FMN、FAD 和 NADPH。Mj NOS mRNA 在斑节对虾的脑、鳃、肠、胸神经节和睪丸中持续表达。当副溶血弧菌注入斑节对虾时，Mj NOS 在脑、鳃、心脏、淋巴器官、肠和胸神经节中表达。注射副溶血弧菌后 12 h 时，鳃中的 Mj NOS 表达达到峰值，24 h 后降至正常水平。
