中国动物检疫2011，Vol.28Issue(9)：27-31,5.
Ⅰ类新城疫病毒NDV08-004株微型基因组的构建及其功能鉴定
Construction and Identification of Mini-genome of Class Ⅰ Newcastle Disease Virus NDV08-004 Strain
摘要
Abstract
Objective To construct the mini-genome system of Class I Newcastle disease virus NDV08-004 strain, providing a foundation of reverse rescue system of the virus. M~aod The mini-genome plasmid, named as pLGR-03, containing the 3＇ and 5＇ terminal regions of class I Newcastle disease virus NDV08-004 isolate linking eGFP as the reported gene in the order of 3＇ Leader-eGFP-5＇ Traile were constructed by over-lap extension （SOE）, and then sub-cloned into transcription vector pOLTV5. The pLGR-03 was transfected into BSR T7/5 cells, infected with NDV08-004 as the helper virus, to identify the function of the mini-genome. Three helper plasmids, pCI-NP, pCI-P and pCI-L,were constructed by sub-cloning the complete ORFs of N-P,P,and L gene into pCI-neo vector respectively. Co-transfections of three helper plasmids together with pLGR-03 were conducted to evaluate the fimction of the helper plasmids. The reporter gene could be expressed after it was tmnsfected into the BSR T7/5 cells infected with helper virus NDV08-004. The BSR T7/5 cell transfected by pLGR-03 and three helper plasmids indicated the eGFP expressed as expected. Concision The work mentioned above will accelerate greatly the rescue of class I NDV and other relative research.关键词
Ⅰ类新城疫病毒/辅助性质粒/微型基因组/病毒拯救Key words
Newcastle disease virus （NDV）/class I/help-plasmid/mini-genome/rescue of virus分类
农业科技引用本文复制引用
陈云霞,管峰,赵云玲,郑东霞,张维,刘华雷,王志亮..Ⅰ类新城疫病毒NDV08-004株微型基因组的构建及其功能鉴定[J].中国动物检疫,2011,28(9):27-31,5.基金项目
国家自然科学基金（30901079) （）
