Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (10): 2094-2102.doi: 10.3864/j.issn.0578-1752.2013.10.015

• ANIMAL SCIENCE·RESOURCE INSECT • Previous Articles     Next Articles

Molecular Cloning and Preliminary Functional Analysis of Domains of Duck Retinoic Acid Inducible Gene I

 CHEN  Yang, HUANG  Zheng-Yang, ZHANG  Yang, LI  Xin-Yu, ZHEN  Ting, WU  Ning-Zhao, XU  Qi, CHEN  Guo-Hong   

  1. 1.College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, Jiangsu
    2.College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045
  • Received:2012-09-28 Online:2013-05-15 Published:2013-02-27

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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, Mx1 and PKR mRNA were detected by real-time PCR.【Result】The whole-length of duRIG-I CDS was 2 802 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.

Key words: duck , RIG-I gene , cloning , RLR signal pathway

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