靶向GP5基因脱氧核酶抑制PRRSV在MARC-145细胞中的复制  

Inhibition of PRRSV replication by DNAzymes targeted to matrix protein GP5 gene in MARC-145 cells

作　　者：宋德武[1] 袁洁[2] 李鑫[1] 郭育培[1] 杨建新[1] 宣华[2] 王贵平[2] 丁壮[1] 

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510640

出　　处：《中国兽医学报》2011年第12期1695-1701,共7页Chinese Journal of Veterinary Science

基　　金：广东省科技攻关重大项目(2008A020100020);吉林大学研究生创新基金资助项目(20111077)

摘　　要：针对猪繁殖与呼吸综合征病毒(PRRSV)JL/07/SW株GP5基因设计了4个脱氧核酶,经体外切割试验筛选得到具有体外切割活性的脱氧核酶,并通过RT-PCR、TCID50、CPE、间接免疫荧光(IFA)检测具有切割活性的脱氧核酶抑制PRRSV复制效果进行评价。结果针对GP5基因的Dz-GP5-10抑制效率最高可达到82.4%,Dz-GP5-12也表现良好的抑制效果,表明GP5基因的这2个位点可能是PRRSV复制所必需的。本研究为PRRSV复制及基因组功能研究、抗病毒药物开发和转基因动物研究奠定了基础。Four DNAzymes targeting the porcine reproductive and respiratory syndrome virus（PRRSV） protein GP5 gene were designed.The active DNAzymes were derived by cleavage experiments in vitro.Marc145 cells were infected with PRRSV 6 h after the active DNAzymes were delivered,efficiency of DNAzymes was assayed by Real-time RT-PCR,TCID50,CPE and indirect immunofluorescence assay（IFA）.This experiment successfully derived the active DNAzyme inhibition of PRRSV replication procedure in MARC-145 cells and showed that the two interference target site may be necessary for PRRSV replication.In future studies,a combination of DNAzyme targeting the GP5 gene may be used as a tool to study PRRSV replication,antiviral therapy and transgenic animals.

关 键 词：PRRSV 脱氧核酶 GP5基因 

分 类 号：S852.65[农业科学—基础兽医学]