猪维甲酸诱导基因I的克隆及其在诱导I型干扰素中的作用

畜牧兽医学报 ›› 2010, Vol. 41 ›› Issue (2): 135-140.doi:

猪维甲酸诱导基因I的克隆及其在诱导I型干扰素中的作用

王荡^1，2，方六荣^1，2 ，梅小伟^1，2 ，谢立兰^1，2 ，陈焕春^1，2 ，肖少波^1，2*

1.华中农业大学动物医学院动物病毒室，武汉 430070；2.华中农业大学农业微生物学国家重点实验室，武汉 430070

收稿日期:2009-04-13 修回日期:1900-01-01 出版日期:2010-02-24 发布日期:2010-02-24
通讯作者: 肖少波

Molecular Cloning and Function Study on Induction of Type I Interferon of Porcine Retinoblastomainhibiting Gene I

WANG Dang ^{1 ，2} ，FANG Liurong^{1 ，2} ，MEI Xiaowei^{1 ，2} ，XIE Lilan ^{1 ，2} ，CHEN Huanchun ^{1 ，2} ，XIAO Shaobo^{1 ，2} *

1.Laboratory of Animal Virology， College of Veterinary Medicine， Huazhong Agricultural University，Wuhan 430070，China; 2.National Key Laboratory of Agriculture Microbiology，Huazhong Agricultural University，Wuhan 430070，China

Received:2009-04-13 Revised:1900-01-01 Online:2010-02-24 Published:2010-02-24
Contact: XIAO Shaobo

摘要/Abstract

摘要： 本研究旨在探讨猪维甲酸诱导基因I (RIGI)的结构特征及其在I型干扰素信号通路中的作用。从猪外周血单核细胞中克隆猪RIGI全长cDNA，进一步构建猪RIGI全长及不同区域缺失突变体的真核表达载体，通过IFNβ、IRF3和NFκB的荧光素酶报告系统分析猪RIGI在诱导I型干扰素中的作用。结果表明，猪RIGI开放读码框全长为2 832 bp，编码943个氨基酸，与鸭嘴兽、大鼠、小鼠、猴、黑猩猩、人、马和牛RIGI相应序列的同源性为53.2%～83.2%。猪RIGI超表达能显著激活转录因子IRF3和NFκB，并诱导IFNβ的产生。缺失猪RIGI的 CARD 区不仅不能激活下游信号，而且还负调控poly(I:C)诱导IFNβ的能力。结果提示RIGI是猪天然免疫系统中的一个模式识别受体，在I型干扰素诱导的信号通路中具有重要作用。

Abstract: This experiment was conducted to study the structural characterization of porcine RIGI (poRIGI) and its role in type I interferon signal pathway. The full length cDNA of poRIGI was amplified from porcine peripheral blood mononuclear cells (PBMCs) by RTPCR. The eukaryotic expression vector of poRIGI fulllength and the mutated poRIGI were constructed, respectively. The role of poRIGI in the induction of type I interferon was investigated by using IFNβ, IRF3 and NFκB luciferase reporter system. The results showed that the open reading frame of poRIGI is 2 832 bp encoding 943 amino acids. The putative poRIGI protein exhibited identity with the corresponding sequences of platypus, rat, mouse, monkey, chimpanzee, human, horse and cattle ranging from 53.2% to 83.2%. Overexpression of the fulllength poRIGI significantly induced the expression of IFNβ by activating transcription factors NFκB and IRF3. The mutant CARD lacking poRIG1 was not capable of activating downstream signals and inhibited poly(I:C)mediated IFNβ production. These results indicate that poRIGI is a pattern recognition receptor in innate immune system of pig and plays an important role in the induction of type I interferon.

王荡;;方六荣;;梅小伟;;谢立兰;;陈焕春;;肖少波;. 猪维甲酸诱导基因I的克隆及其在诱导I型干扰素中的作用[J]. 畜牧兽医学报, 2010, 41(2): 135-140.

WANG Dang ;;FANG Liurong ;;MEI Xiaowei ;;XIE Lilan ;;CHEN Huanchun ;;XIAO Shaobo ;. Molecular Cloning and Function Study on Induction of Type I Interferon of Porcine Retinoblastomainhibiting Gene I[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2010, 41(2): 135-140.

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