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Issue:ISSN 1000-7083
          CN 51-1193/Q
Director:Sichuan Association for Science and Technology
Sponsored by:Sichuan Society of Zoologists; Chengdu Giant Panda Breeding Research Foundation; Sichuan Association of Wildlife Conservation; Sichuan University
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Your Position :Home->Past Journals Catalog->2011 Vol.30 No.4

Cloning and Sequence Analysis of the cDNA and Promoter of Grass Carp Inducible Nitric Oxide Synthase
Author of the article:WANG Xin-yan, ZHAO Tai-qiang, GUO Jia-cong, YANG Mu, ZHOU Hong*
Author's Workplace:(College of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu 610054, China)
Key Words:iNOS;cDNA sequence;promoter sequence;sequence analysis
Abstract:The full length cDNA of grass carp iNOS was isolated from LPS-stimulated grass carp head kidney leucocytes by using PCR amplification coupling to 5’- and 3’- RACE. This cDNA was 4286 bp in length which encodes a protein with 1080 amino acid residues. Amino acid alignment showed that the identities between grass carp iNOS and goldfish iNOS, zebrafish iNOS2b and common carp iNOS were all more than 80%. The cofactor-binding sites for heme, tetrahydrobiopterin, calmodulin, FMN, FAD pyrophosphate, FAD isoalloxazine, NADPH ribose, NADPH adenine and NADPH are also highly conserved in these sequences. Phylogenetic analysis confirmed that this newly cloned iNOS is a member of inducible NOS. The promoter sequence of grass carp iNOS with 1978 bp was also obtained by using the Genome Walker Universal kit. Multiple transcription factors including GR, AP1, C/EBP, ER, YY1, IRF1 and IL-6 REBP binding sites were revealed by using transcription element search system in this study.
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