新城疫病毒耐热弱毒TS09-C株反向遗传操作系统的建立  

Establishing Reverse Genetic System of Thermostable Avirulent Newcastle Disease Virus Strain TS09-C

作　　者：温国元[1] 郭静[1,2] 商雨[1,2] 陈晨[1,2] 王红琳[1] 罗青平[1] 张蓉蓉[1] 王宏才[1] 张腾飞[1] 杨峻[1] 程国富[2] 邵华斌[1] 

机构地区：[1]湖北省农业科学院畜牧兽医研究所,武汉430064 [2]华中农业大学动物医学院,武汉430070

出　　处：《湖北农业科学》2014年第24期6075-6078,6081,共5页Hubei Agricultural Sciences

基　　金：现代农业产业技术体系建设专项资金(CARS-42-G11);公益性行业(农业)科研专项(201303033);湖北省农业科技创新中心资助项目(2011-620-001-003);动物胚胎及分子育种湖北省重点实验室开放课题项目(2012ZD151)

摘　　要：为了建立新城疫病毒(NDV)耐热弱毒株的反向遗传操作平台,以NDV TS09-C耐热弱毒株为研究对象,构建了TS09-C株的全长c DNA克隆质粒和3个分别表达NP、P和L的辅助质粒。通过4个质粒共转染预感染有表达T7 RNA聚合酶的痘病毒的BHK-21细胞,拯救出了重组病毒r TS09-C株。生物学特性试验结果表明,重组病毒r TS09-C株具有与亲本株相似的耐热、弱毒和细胞增殖特性。NDV耐热毒株反向遗传操作系统的建立为耐热活疫苗载体的研究,进一步研发新型多联(多价)耐热禽用活疫苗提供了技术平台,也为耐热机理的研究打下基础。To establish the reverse genetic system of the thermostable avirulent NDV strain TS09-C, the full-length cDNA clone plasmid, along with three supporting plasmids expressing the NP, P and L gene, was constructed and co-transfected into the BHK-21 cells pre-inoculated with vTF7-3. The recombinant virus rTS09-C was rescued from BHK-21 cell. The NDV rTS09-C was identified by IFA and electron microscope. The results of the biological characterization confirmed that the rTS09-C virus had the same thermostability, low pathogenicity and cell growth curve as its parental strain TS09-C. It is indicated that the reverse genetic system of the thermostable avirulent NDV was successfully established, which was valuable for further developing multi-valent avian thermostable vaccines and studying the NDV thermostable mechanisms.

关 键 词：新城疫病毒(NDV) 耐热 反向遗传操作 

分 类 号：S852.659.5[农业科学—基础兽医学]