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

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.