石斑鱼一氧化氮合酶cDNA的分子克隆及序列分析  

Molecular cloning and sequence analysis of nitric oxide synthase gene from Epinephelus coioides

作　　者：江湧[1] 谢骏[1] 李文笙[1] 林浩然[1] 吴淑勤[2] 

机构地区：[1]中山大学水生经济动物研究所暨广东省水生经济动物重点实验室,广东广州510275 [2]中国水产科学研究院珠江水产研究所,广东广州510380

出　　处：《水产学报》2003年第4期381-385,共5页Journal of Fisheries of China

基　　金：国家海洋863项目资助(2001AA621010;2001AA622050)

摘　　要：Nitric oxide (NO) is a highly reactive, labile gas produced by the enzymatic conversion of L-arginine by nitric oxide synthases NOS. In mammals, NO mediates multiple physiological processes from cardiovascular control to neural transmission. Three NOS isoforms neuronal, inducible and endothelial have been cloned, sequenced and characterized from several mammalian species. nNOS isoforms were characterized from adult grouper (Epinephelus coioides) by using RT-PCR with degenerate oligonucleotide primers designed against a portion of the mammalian NOS gene that codes for the calmodulin -binding region, this region was chosen because it is highly conserved among NOS sequences to date and is a functionally important region of NOS proteins. A partial gene sequence of 377 bp corresponding to mammalian nNOS is obtained. This sequence showed 82%-83%, 85%-93% homogeneity with that of mammalian and the other fish respectively. The deduced amino acid of E.coioides shows high identity with that of mammalian (92%-93%), and the other fish (93%-99%). Phylogenetic analysis of these sequences confirms the conserved nature of NOS, particularly of the calmodulin-binding domains.Nitric oxide (NO) is a highly reactive, labile gas produced by the enzymatic conversion of L-arginine by nitric oxide synthases NOS. In mammals, NO mediates multiple physiological processes from cardiovascular control to neural transmission. Three NOS isoforms neuronal, inducible and endothelial have been cloned, sequenced and characterized from several mammalian species. nNOS isoforms were characterized from adult grouper (Epinephelus coioides) by using RT-PCR with degenerate oligonucleotide primers designed against a portion of the mammalian NOS gene that codes for the calmodulin -binding region, this region was chosen because it is highly conserved among NOS sequences to date and is a functionally important region of NOS proteins. A partial gene sequence of 377 bp corresponding to mammalian nNOS is obtained. This sequence showed 82%-83%, 85%-93% homogeneity with that of mammalian and the other fish respectively. The deduced amino acid of E.coioides shows high identity with that of mammalian (92%-93%), and the other fish (93%-99%). Phylogenetic analysis of these sequences confirms the conserved nature of NOS, particularly of the calmodulin-binding domains.

关 键 词：石斑鱼 一氧化氮合酶 CDNA 分子克隆 序列分析 钙调蛋白结合区 

分 类 号：S965.334[农业科学—水产养殖]