渔业科学进展2012，Vol.33Issue(5)：39-47,9.
半滑舌鳎IRF-7基因的全长cDNA克隆及表达分析
Molecular cloning and tissue expression of IRF-7 gene of Cynoglossus semilaevis
摘要
Abstract
The full-length cDNA sequence of interferon regulatory factor 7 (IRF-7) of tongue sole Cynoglossus semilaevis was cloned by rapid amplification cDNA end (RACE) method. The full-length cDNA of C. semilaevis IRF-7 (CsIRF-7) is 1 957bp, containing a 48 bp 5'- untranslated region (5'-UTR) , 1 302 bp open reading frame (ORF), and a 607 bp 3 -UTR. The putative amino acid sequence of CsIRF-7 contains 433 amino acid residues. Through protein sequence alignment of IRF-7 in vertebrates,we found that the IRF-7 contains three completely conserved residues across all species, which are DNA-binding domain (DBD), IRF associated domain (IAD) and serine-rich domain (SRD). In contrast to IRF-7 of vertebrates other than fish, the DBD which is highly conserved has five tryptophan repeats, while CsIRF-7 and other fish IRF-7s have only four tryptophans with the second tryptophan residue missing. The deduced amino acid sequence of CsIRF-7 exhibited a high-level of homology with other fish, and shared the highest similarity (83. 8%) and identity (73. 6%) with the IRF-7 of Paralichthys olivaceus. The CsIRF-7 shared a low-level of homology with mammalia and aves. Phylogenetic analysis revealed that the C. semilaevis IRF-7 was clustered into IRF-7 subgroup with fish IRF-7 and formed the IRF-3 subfamily with IRF-3 group and other vertebrates IRF-7. Quantitative real-time method was used to detect the CsIRF-7 mRNA expression in different tissues. The results showed that CsIRF-7 was expressed in all tested tissues including liver, kidney, spleen, gill, intestine, blood cells, and heart. The high-level expression of CsIRF-7 is detected in the intestine, heart, kidney, and spleen, while a lower-level expression in the liver, gill, and blood cells. These results will be helpful for further studies on functions of antiviral and antibacterial of CsIRF-7 gene.关键词
半滑舌鳎/干扰素调节因子7/Poly Ⅰ:C/基因克隆/表达分析Key words
Cynoglossus semilaevis/Interferon regulatory factor-7/Poly I:C Gene cloning/Expression分类
农业科技引用本文复制引用
刘万建,史成银,宋展,耿伟光,李赟,黄倢..半滑舌鳎IRF-7基因的全长cDNA克隆及表达分析[J].渔业科学进展,2012,33(5):39-47,9.基金项目
中国水产科学研究院基本科研业务费(2012A0501)和国家重点基础研究发展计划课题(2006CB101802)共同资助 （2012A0501）
