摘要
用SrfⅠ限制性内切酶将构建的猪瘟病毒兔化弱毒株(C 株)全长cDNA线性化,体外转录获得其基因组RNA,利用脂质体将基因组RNA转染至SK 6细胞系,连续传代,收集5代以后培养细胞,采用RT PCR、荧光抗体直接法、夹心ELISA进行鉴定。结果表明,所构建的全长cDNA具有感染性。猪瘟病毒C 株全长感染性cDNA的成功构建,为在分子水平上进一步研究中国猪瘟病毒兔化弱毒株提供了极有价值的研究工具。
Constructed full-length cDNA pGEM-5Zf(+)/F1-7 of CSFV C-strain was linearized by restriction enzyme SrfⅠ,genomic RNA was in vitro transcribed and further transfected into cell line SK-6 by liposomes. Cells were collected after being passaged 5 times continuously,and were used to identify infection by RT-PCR,direct fluorescent antibody,and sandwich ELISA.The results indicated that infectious RNA of CSFV could be obtained from constructed full-length cDNA.Successful construction of infectious full-length cDNA of CSFV C-strain has provided an excellent tool for further study of CSFV C-strain on level of molecular biology.
出处
《病毒学报》
CAS
CSCD
北大核心
2004年第2期143-147,共5页
Chinese Journal of Virology
基金
国家重点基础研究发展规划(No.G19990119)
关键词
猪瘟病毒
兔化弱毒株
CDNA
克隆
感染性
转染
体外转录
classical swine fever virus(CSFV)
C-strain
full-length cDNA
in vitro transcription
transfection
infection identification
