摘要
根据I型马立克病病毒(MDV)强毒GA株基因序列,设计两对引物,用PCR方法扩增出CVI988／Rispens株的US10及其侧翼序列,分别克隆入pUC18载体中。经测序检测正确后,进一步插入含CMV启动子的绿色荧光蛋白 (EGFP)基因表达盒,获得了含EGFP报告基因的转移载体质粒pPUC18-US10-EGFP。通过同源重组,成功地筛选出表达EGFP的重组病毒rCVI988-EGFP,经传代证明重组rCVI988-EGFP在感染的CEF细胞中能稳定表达EGFP。结果表明:构建的重组转移载体质粒正确,US10是MDV复制非必需片段,为进一步利用US10区构建重组MDV多价基因工程疫苗奠定了基础。
The US10 and its flanks of CV1988/Rispens strain were amplified by PCR with primers designated according to the sequence of “virulent” GA strain of Marek＇s disease virus （MDV） serotype I and subcloned into pUC18 plasmid, named as pUC-US10. The expression cassette including EGFP gene controlled by CMV promoter and the enhancer was then cloned into pUC-US10 vector as a marker of the transfer vector pUC18-US10-EGFP. A stable recombinant Marek＇s disease virus （rMDV） expressing EGFP was obtained by homologous recombination with virus of CVI988/Rispens strain in CEF. These results proved that the US10 of MDV is one of non-essential sites in the viral genome for viral growth in vitro. It facilities the construction of recombinant MDV expressing foreign genes in future.
出处
《扬州大学学报（农业与生命科学版）》
CAS
CSCD
北大核心
2005年第4期1-4,共4页
Journal of Yangzhou University：Agricultural and Life Science Edition
基金
国家高技术研究发展计划项目(863-2002AA245051)全国博士学位论文作者专项资金(200256)
