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3个RNA干扰靶位,构建shRNA表达质粒;将转染干扰质粒6 h后的MARC-145细胞接种病毒,并通过Real-time RT-PCR、TCID50、CPE、间接免疫荧光检测(IFA)对所设计的shRNA表达质粒的干扰效果进行评价;结果表明,构建的干扰质粒可以高效抑制PRRSV在MARC-145细胞中的复制,说明GP5基因的这3个干扰靶位可能是PRRSV复制所必需的。本试验为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。
Three plasmids that express three shRNAs targeting the porcine reproductive and respiratory syndrome virus（PRRSV） protein GP5 gene were designed.Marc145 cells were infected with PRRSV 6 h after the shRNA-expressing plasmids were delivered,efficiency of RNA interfering was assayed by Real-time RT-PCR,TCID50,CPE and indirect immunofluorescence assay（IFA）.This paper successfully revealed the shRNA expressing plasmids RNAi phenomenon of PRRSV replication procedure in MARC-145 cells and showed that the three interference target site may be necessary for PRRSV replication.In future studies,a combination of siRNAs targeting the GP5 gene may be used as a tool to study PRRSV replication,antiviral therapy and transgenic animals.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2010年第8期1013-1017,共5页
Chinese Journal of Veterinary Science
基金
广东省科技攻关重大项目(2008A020100020)
