中国农业科学2013，Vol.46Issue(10)：2094-2102,9.DOI:10.3864/j.issn.0578-1752.2013.10.015
鸭维甲酸诱导基因Ⅰ克隆及其结构域功能分析
Molecular Cloning and Preliminary Functional Analysis of Domains of Duck Retinoic Acid Inducible Gene Ⅰ
摘要
Abstract
[Objective] Duck RIG-Ⅰ (duRIG-Ⅰ) gene was cloned and the functions of its different domains were analyzed preliminarily.[Method] The CDS of duRIG-Ⅰ gene was cloned on the basis of the sequence submitted to GenBank with RT-PCR and was analyzed by bioinformatics.The eukaryotic expression vectors of N-terminal,C-terminal and whole-length of duRIG-Ⅰ gene with 6*his tags were constructed to transfect DF1,and then the transcription and expression of the three recombinant plasmids in cells were detected via RT-PCR and indirect immunofluorescent assay,respectively.Meanwhile,the expressions of chicken IFN-β,Mxl and PKR mRNA were detected by real-time PCR.[Result] The whole-length of duRIG-Ⅰ CDS was 2802 bp encoding 933 amino acids.All the recombinant protein of duRIG-Ⅰ could express normally in DF1.The results of RT-qPCR indicated that CARDs significantly up-regulated the mRNA level of IFN-β,Mx1 and PKR genes.[Conclusion] The various domain fragments of duRIG-Ⅰ express normally in DF1.The N-terminal of duRIG-Ⅰ plays a vital role in regulating the expression of downstream genes of RLR antiviral signal pathway.关键词
鸭/RIG-Ⅰ基因/克隆/RLR通路Key words
duck/ RIG-Ⅰ gene/ cloning/ RLR signal pathway引用本文复制引用
陈阳,黄正洋,张扬,李欣钰,甄霆,吴宁昭,徐琪,陈国宏..鸭维甲酸诱导基因Ⅰ克隆及其结构域功能分析[J].中国农业科学,2013,46(10):2094-2102,9.基金项目
现代农业产业技术体系建设专项(CARS-43-3)、江苏高校优势学科建设工程资助项目(苏政办发[2011]137号) （CARS-43-3）
