猪RIG-I真核表达载体的构建及其抗口蹄疫病毒作用研究

doi:10.11843/j.issn.0366-6964.2015.04.013

ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2015, Vol. 46 ›› Issue (4): 600-607.doi: 10.11843/j.issn.0366-6964.2015.04.013

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Construction of Porcine RIG-I Eukaryotic Expressing Plasmid and Its Antiviral Effects Research against Foot and mouth Disease Virus

WANG Guo-qing¹，ZHU Zi-xiang²，CAO Wei-jun ²，YANG Fan ²，MAO Ruo-qing ²，LI Dan ²，LIU Lei^1*，ZHENG Hai-xue ^2*

(1.College of Veterinary Medicine，Gansu Agricultural University，Lanzhou 730070，China；2.National Foot and Mouth Disease Reference Laboratory，State Key Laboratory of Veterinary Etiological Biology，Lanzhou Veterinary Research Institute，Chinese Academy of Agricultural Sciences，Lanzhou 730046，China)

Received:2014-07-28 Online:2015-04-23 Published:2015-04-23

Abstract

Abstract:

To investigate whether pattern recognition receptor RIG-I has antiviral role against foot-and-mouth disease virus (FMDV)，we extracted porcine cellular RNA from PK15 cells，and the complete CDS region of RIG-I was obtained by using two-segmental amplification and fusion PCR methods.Subsequently，the eukaryotic expressing plasmid was constructed.The expression of the plasmid was confirmed by Western blotting and indirect fluorescence assay (IFA)，which showed RIG-I was successfully expressed.The IFA result indicated porcine RIG-I protein was located in the cellular cytoplasm.The results of infection assay suggested that FMDV infection induced the upregulation of RIG-I expression，which implied the potential connection between RIG-I and FMDV.Over-expression assay confirmed that RIG-I inhibited FMDV replication，and downregulation of RIG-I significantly promoted FMDV replication.These results indicated that RIG-I had antiviral effect against FMDV.In conclusion，this study provided references for further research on antiviral mechanism of RIG-I against FMDV.It also paves ways for the research of antiviral mechanism of innate immune system after FMDV infection.

CLC Number:

S582.659.6

WANG Guo-qing,ZHU Zi-xiang,CAO Wei-jun,YANG Fan,MAO Ruo-qing,LI Dan,LIU Lei,ZHENG Hai-xue . Construction of Porcine RIG-I Eukaryotic Expressing Plasmid and Its Antiviral Effects Research against Foot and mouth Disease Virus[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(4): 600-607.

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Construction of Porcine RIG-I Eukaryotic Expressing Plasmid and Its Antiviral Effects Research against Foot and mouth Disease Virus

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