RNAi靶向N基因抑制猪繁殖与呼吸综合征病毒的复制  

Inhibition of porcine reproductive and respiratory syndrome virus (PRRSV) replication by RNA interference targeted to N gene

作　　者：杨闽楠[1] 李志杰[1] 丁壮[1] 张泉鹏[1] 宣华[2] 王贵平[2] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062 [2]广东省农业科学院兽医研究所,广东广州510000

出　　处：《中国兽医学报》2010年第12期1571-1575,1579,共6页Chinese Journal of Veterinary Science

基　　金：广东省科技攻关重大资助项目(2008A020100020)

摘　　要：根据编码猪繁殖与呼吸综合征病毒(PRRSV)核衣壳蛋白(N)基因的序列,设计3个干扰靶位(N12、N23、N26),构建siRNA表达载体,转染Marc-145细胞后接毒,测定病毒TCID50、CPE,并利用实时荧光定量PCR及间接免疫荧光检测病毒在Marc-145细胞上的复制情况。结果表明,利用RNAi技术靶向N基因,其中N12干扰靶位可以高效抑制PRRSV的增殖,证实N基因可能是病毒复制所必需的结构基因,所选的干扰靶位是病毒复制的关键性位点。According to the subsequence of nucleocapsid protein of porcine reproductive and respiratory syndrome virus（PRRSV）,three interference target sites（N12,N23,N26） were designed,and constructed siRNA expression vectors.Then the recombine RNAi vector was transfected into Marc-145 cells which was infected by PRRSV after.The condition of virus replication in Marc-145 cells was detected by TCID50,CPE,real-time quantitative PCR and indirect immunofluorescence.Experimental results showed that RNAi technology can inhibit PRRSV proliferation by targeted to N12 gene efficiently,and confirm that N gene may be the imperative structural gene for viral replication.Meanwhile,the selected interference target site is a key site of viral replication.

关 键 词：猪繁殖与呼吸综合征病毒 N基因 RNA干扰 

分 类 号：S852.65[农业科学—基础兽医学]