Molecular Cloning and Characterization of the Promoter Region of the Inducible Nitric Oxide Synthase Gene of the Rat

Jin-kun Wen^1*, Mei Han¹, and Dian-wu Liu²

¹Department of Biochemistry and ²Department of Medical Microbiology, Hebei Medical University, Shijiazhuang 050017, People's Republic of China; fax: 0086-311-6048177; E-mail: wjk@hebmu.edu.cn

^* To whom correspondence should be addressed.

Received March 15, 2000; Revision received June 15, 2000

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To investigate the mechanism by which inducible nitric oxide synthase (iNOS) within vascular smooth muscle cells (VSMC) is expressed following exposure to proinflammatory cytokines, we cloned the promoter region of the rat iNOS gene by a single specific primer PCR cloning strategy. The 5´-flanking region of the rat iNOS gene contains interferon-gamma (IFN-gamma)- and tumor necrosis factor-alpha (TNF-alpha)-responsive elements and an NF-kappaB-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(s) which bound specifically to the fragment of the rat iNOS gene containing several consensus sequences were produced after VSMC were treated with interleukin 1 (IL-1) and IFN-gamma.

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KEY WORDS: inducible nitric oxide synthase, promoter region, cloning, sequence analysis, rat

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