摘要
本研究旨在建立猪脑心肌炎病毒(EMCV)的感染性克隆技术。利用RT-PCR分3段扩增出脑心肌炎病毒BJC3株的全基因组cDNA,依次克隆至低拷贝质粒pWSK29,构建出全长质粒pWSKBJC3/w,经体外转录和转染BHK-21细胞拯救病毒。结果表明,构建的全长cDNA克隆具有感染性,在BHK-21细胞上可拯救出病毒。拯救病毒(命名为rVBJC3W)在BHK-21细胞上的生长特性与其亲本病毒BJC3一致,并保持了对小鼠的致病性。本研究成功构建了中国第1株猪脑心肌炎病毒的感染性克隆,为深入研究其分子致病机制提供了必要的工具。
The objective of this study was to construct infectious clone of encephalomyocarditis virus（EMCV）.The genomic cDNA of EMCV BJC3 was amplified by three overlapped segments using RT-PCR,and cloned into low-copy plasmid pWSK29 to construct full-length cDNA clone pWSKBJC3/w.The pWSKBJC3/w was in vitro transcribed and transfected into BHK-21 cells to rescue the virus.The results showed that the full-length cDNA clone was infectious and the virus could be rescued on BHK-21 cells.The rescued virus designated rVBJC3W was identified by RT-PCR and indirect immunofluorescence assay（IFA）.The rescued virus had similar growth characteristics with its parental virus BJC3 and remained the pathogenicity for mice.Our results indicated that the first infectious cDNA clone of EMCV in China was successfully established and provided an essential tool for investigating the molecular basis of pathogenicity of EMCV.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2009年第9期1350-1357,共8页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家自然科学基金重点项目(30530550)
关键词
脑心肌炎病毒
感染性克隆
病毒拯救
生长特性
致病性
encephalomyocarditis virus （EMCV）
infectious eDNA clone
virus rescue
growthcharacteristics
pathogenicity
