NDVsdbz-s99分离株的生物学特性及其F基因的克隆与序列分析

The Biological Characteristics of NDV sbdz-s99 Isolate and Cloning and Sequencing of Its F Gene

对山东省新城疫分离株sdbz_s99的生物学特性进行了鉴定 ;并以其基因组RNA为模板 ,经RT_PCR扩增出含有F蛋白裂解位点的基因片段。克隆到载体pMD18_T后 ,转化大肠杆菌 ,通过双脱氧末端终止法进行核苷酸序列分析。结果表明 ,所克隆片段大小为 36 4bp。经Blastn分析表明与已发表的NDV各代表株同源性为 83.0 %～ 98.0 % ,特别是与目前广泛应用的弱毒疫苗LaSota株同源性较低 ,仅为 83.0 %。利用MegAlign软件 ,以前 36 4bp绘制了NDV的系统发育进化树 ,其结果表明该分离株为基因VII型 ,说明山东省也有基因VII型新城疫的流行。

The Biological Characteristics of NDV sbdz_s99 Isolate were identified.Gene fragment with F protein splitting site was amplified by RT_PCR using its genomic RNA as template,and cloned into pMD18_T and then E.coli were transformed.The suquencing was performed by dideoxy method.The result of the sequencing demonstrated that the cloned fragment is 364bp.The homogeneity among this isolate and those representative NDV strains reported is 83.0%～98.0% by Blasten analysis,but the homogeneity between the isolate and LaSota strain is only 83.0%.The NDV systemic evolution tree was depicted with the 364bp sequence by MegAlign software.The result showed that the isolate is gene type VII and the ND caused by gene type VII also prevail in Shandong province.