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Position: Home > Articles > Inhibition of PRRSV replication by DNAzymes targeted to matrix protein GP5 gene in MARC-145 cells Chinese Journal of Veterinary Science 2011,31 (12) 1695-1701

靶向GP5基因脱氧核酶抑制PRRSV在MARC-145细胞中的复制

作  者:
宋德武;袁洁;李鑫;郭育培;杨建新;宣华;王贵平;丁壮
单  位:
吉林大学畜牧兽医学院;广东省农业科学院兽医研究所
关键词:
PRRSV;脱氧核酶;GP5基因
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。
译  名:
Inhibition of PRRSV replication by DNAzymes targeted to matrix protein GP5 gene in MARC-145 cells
作  者:
SONG De-wu1,YUAN Jie2,LI Xin1,GUO Yu-pei1,YANG Jian-xin1,XUAN Hua2,WANG Gui-ping2,DING Zhuang1(1.College of Animal Science and Veterinary Medicine,Jilin University,Changchun 130062,China;2.Institute of Veterinary Medicine,Guangdong Academy of Agricultural Science,Guangzhou 510640,China)
关键词:
PRRSV;DNAzyme;GP5 gene
摘  要:
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.

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