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Cloning and characterization of interferon stimulated genes Viperin and ISG15,and their promoters from snakehead Channa argus

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Progress in Natural Science:Materials International 2007年第12期17卷 1425-1435页

作者： jia weizhang1.2, zhou xiuxia1.2, huang rong1.2 and guo qionglin1.1. institute of hydrobiololgy, chinese academy of sciences, wuhan 430072, china 2. Graduate University of chinese academy of sciences, Beijing 100039, china) institute of Hydrobiology Chinese Academy of Sciences Wuhan 430072 China Graduate University Chinese Academy of Sciences Beijing 100039 China

By suppression subtractive hybridization, rapid amplification of cDNA ends and gene walking methods, interferon stimulated genes (ISGs), Viperin and ISG15, and their promoters have been cloned and characterized from snakehead Channa argus. The Viperin cDNA was found to be 1474 nt and contain an open reading frame (ORF) of 1059 nt that translates into a putative peptide of 352 amino acid (aa). The putative peptide of Viperin shows high identity to that in teleosts and mammals except for the N-terminal 70 aa. The ISG15 cDNA was found to be 758 nt and contain an ORF of 468 nt that translates into a putative peptide of 155 aa. The putative peptide of ISG15 is composed of two tandem repeats of ubiquitin-like (UBL) domains, and a canonical conjugation motif (LRGG) at C-terminal. Viperin and ISG15 promoter regions were characterized by the presence of interferon stimulating response elements (ISRE) and γ-IFN activation sites (GAS). ISRE is a feature of IFN-induced gene promoter and partially overlaps interferon regulatory factor (IRF) 1 and IRF2 recognition sites. GAS is responsible for the γ-IFN mediated transcription. One conserved site for NF-κB was found in the promoter region of Viperin. This is the first report of conservative binding motif for NF-κB in accordance with the consensus sequence (GGGRNNYYCC) among teleost ISG promoters. Moreover, there were also TATA, CAAT and Sp1 transcription factor sites in Viperin and ISG15 promoters. In 5′ untranslated region (UTR), snakehead ISG15 gene contains a single intron, which differs from Viperin gene. The transcripts of Vipeirn and ISG15 mRNA were mainly expressed in head kidney, posterior kidney, spleen and gill. The expression levels in liver were found to increase obviously in response to induction by IFN-inducer poly I∶C.

关键词： interferon interferon stimulated gene (ISG) Viperin ISG15 snakehead (Channa argus).

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