大鼠诱导型一氧化氮合酶基因转录调控区的克隆与鉴定

采用单特异引物ＰＣＲ克隆法，得到大鼠诱导型一氧化氮合酶（ｉＮＯＳ）基因转录调控区ＤＮＡ片段．核酸序列分析证实，大鼠ｉＮＯＳ基因的５′－侧翼区含有ＩＦＮ－γ和ＴＮＦ－α应答元件及ＮＦ－κＢ结合位点的保守序列．这些保守序列的位置及排列显著区别于人和小鼠的ｉＮＯＳ基因．电泳迁移率改变分析（ＥＭＳＡ）表明，ＶＳＭＣ受ＩＬ－１和ＩＦＮ－γ刺激后，细胞核内产生某种可与ｉＮＯＳ基因５′－侧翼区特异结合的核蛋白因子．

Molecular Cloning and Characterization of Upper Regulation Region of Rat Inducible Nitric Oxide Synthase Gene

To investigate the mechanism by which inducible nitric oxide synthase (iNOS) within vascular smooth muscle cells (VSMC) is expressed following exposure to proinflammatory cytokines.The promoter region of rat iNOS gene was cloned by a single specific primer PCR cloning strategy.The 5′ flanking region of the rat iNOS gene contains IFN γ and TNF α responsive elements and NF κ B binding consensus sequences.The position and alignment of these consensus sequences are distinct from those in the mouse iNOS and human iNOS genes.It was shown that some nuclear factor which bound specifically to the fragment of the rat iNOS gene containing several consensus sequences were produced after VSMC was treated with IL 1 and IFN γ.