猪TREX1基因的真核表达及其在JEV感染中的作用研究  

Eukaryotic Expression of Porcine TREXI Gene and Its Effect on Japanese Encephalitis Virus Infection

作　　者：Safdar Anum 李慧 李壮壮 相笑 魏建超[1] 刘珂[1] 邵东华[1] 李蓓蓓[1] 马志永[1] 邱亚峰[1] Safdar Anum;LI Hui;LI Zhuangzhuang;XIANG Xiao;WEI Jianchao;LIU Ke;SHAO Donghua;LI Beibei;MA Zhiyong;QIU Yafeng(Shanghai Veterinary Research Institute,CAAS,Shanghai 200241,China)

机构地区：[1]中国农业科学院上海兽医研究所,上海200241

出　　处：《中国动物传染病学报》2022年第1期1-5,共5页Chinese Journal of Animal Infectious Diseases

基　　金：上海市自然基金(15ZR1449800);国家重点研发计划(2016YFD0500404);中国农业科学院青年英才(CAASQNYC-KYYJ-26)。

摘　　要：为了明确猪TREX1在JEV感染中的作用,本研究利用PCR扩增获得猪TREX1基因的ORF,并将该基因克隆至真核表达载体p3xFlag-CMV-14中,将构建的重组真核表达质粒命名为pFlag-pTREX1,利用lipofectamine 2000将构建成功的pFlag-pTREX1重组质粒瞬时转染至293T细胞中,通过Western blot检测其表达情况,最后在PIEC细胞上,通过猪TREX1基因过表达或沉默TREX1基因的表达,分析其在JEV感染中的作用。结果显示:本研究成功构建了Flag-pTREX1真核表达载体;利用抗Flag抗体可以识别真核表达的Flag-pTREX1融合蛋白,约35 kDa;与转染Flag-vector的样品相比,过表达猪TREX1促进JEV在PIEC上的感染;与NC对照组相比,沉默猪TREX1基因的表达抑制JEV在PIEC上的表达。结果表明,本研究成功构建了猪TREX1基因真核表达载体并初步探索了猪TREX1在JEV感染中的作用,这为进一步研究猪TREX1在猪源病毒中的作用奠定了基础。In order to understand eukaryotic expression characteristics of porcine TREX1(pTREX1)and its role in Japanese encephalitis virus(JEV)infection,a recombinant plasmid pFlag-pTREX1 was constructed by inserting porcine TREX1 gene into p3xFlag-CMV-14 by PCR and transient expression of porcine TREX1(pTREX1)in 293T cell line was determined by Western blot.The role of pTREX1 in JEV infection was determined in PIEC cell line using overexpression or RNA interference(RNAi).The results showed that Flag-pTREX1 was generated and the recombinant protein Flag-pTREX1 of about 35 kDa was specifi cally detected by anti-Flag antibody.In addition,overexpression of pTREX1 in PIEC cell line promoted JEV infection compared with Flag-vector group.Lastly,the knock-down expression of porcine TREX1 gene inhibited JEV infection as compared with NC group.Collectively,our data demonstrate eukaryotic expression characteristics of porcine TREX1(pTREX1)and its role in JEV infection.These data would also provide basis of understanding role of pTREX1 in viruses.

关 键 词：TREX1 真核表达 RNA干扰 抗病毒反应 JEV 

分 类 号：S852.659.6[农业科学—基础兽医学]