鹅源副黏病毒NA-1株反向遗传操作体系的建立

Construction of reverse genetics system of GPMV NA-1 strain

应用cRACE等方法扩增鹅源副黏病毒NA-1株cDNA5′末端和3′末端,分6段扩增得到病毒的结构基因序列,而后连接本室构建的TLH-T转录载体,构建其cDNA全长克隆。在引入分子标签和验证辅助质粒的功能后,4质粒系统共转染VT7细胞系,成功拯救出了具有感染性的鹅副黏病毒。鹅源副黏病毒NA-1株反向遗传操作体系的建立为进一步深入研究该病毒基因组的功能以及新型疫苗的研发奠定了基础。

To establish the rescue system for goose host paramyxovirus NA-1 strain by reverse genetic,we connected correctly the transcription vector with the 3＇-and 5＇-terminal ends and the full-lenth cDNA clong. After validated the genetic markers and the three helper plasmids,four plasmids were cotransfected into VT7 cells expressing T7 RNA polymerase. The typical morphology of the rescued GPMV was detected in the electronic microscope. The result in dicats we have rescued the GPMV NA-1 strain successful. The reverse genetics system provids a basic technical platform for further research on gene function and novel vaccine candidate of GPMV NA-1 strain.