作　　者： ; ; ; ; ;

机构地区： 山东省农业科学院

出　　处： 《农业生物技术学报》 2014年第3期273-279,共7页

摘　　要： 新城疫(newcastle disease,ND)是禽类的两大疫病之一,新城疫病毒(Newcastle disease virus,NDV)是该病的病原体。为了解野禽NDV的分子特性、致病性及其与家禽NDV之间的关系,2011年分离到一株野禽NDV,将其命名为NDV2;通过标准生物学毒力检测证实其为强毒株。参照GenBank发布的NDV全长基因序列设计9对特异性引物,对NDV2进行全基因的序列测定和分析。拼接后序列分析表明,NDV2株的全基因组序列总长为15 192 nt,包含6个开放阅读框,分别编码6种蛋白nucleocapsid protein(NP)、phosphoprotein(P)、matrix protein(M)、fusion protein(F)、haemagglutinin-neuraminidase(HN)和large protein(L)的基因,长度分别为1 753、1 451、1 241、1 792、2 004和6 703 nt(GenBank登录号:KF306265)。与国内多数禽源分离毒株相似,NDV2毒株亦属于ClassⅡ、基因Ⅶ型。综合各个蛋白的同源性比较,NDV2株与近年国内流行的一些基因Ⅶ型毒株如:Egret.GX.11、Duck.WF.00、Goose.GD450.11等的同源性较高,远高于其他地区和其他基因型的毒株,这说明NDV2毒株与国内的野毒株亲缘性较高;与NDV2同源性差距最大的为ClassⅠ的JS10毒株。F基因的裂解位点显示:全基因长度为15 192nt的毒株(包括NDV2毒株在内)在裂解位点处的氨基酸序列为RRQKRF或KRQKRF,为强毒株序列。从基因分型结果来看,NDV2毒株和绝大多数参考毒株F基因和HN基因分型结果是一致的;唯一不同的就是印度鸡源分离株NDV4毒株,F基因分型它属于Ⅱ型,HN基因分型它属于Ⅶ型。分析各个基因的起始序列和终止序列后发现:NDV2株与其它参考NDV毒株的各个基因的起始序列完全相同,说明各基因起始序列保守性较高;不同之处是NDV2毒株F和HN基因的终止序列与La Sota和Sweden95一致,与其他野禽毒株不同。以NDV2人工感染无特定病原(Spicific pathogenfree,SPF)鸡(Gallus gallus)、鸭(Anas platyrhynchos platyrhynchos Linnaeus.)和鸽(Rupestris Pallas),并且每个攻� Newcastle diseases （ND） is one of the two serious diseases of poultry, Newcastle diseases virus （NDV） is the pathogen of the disease. To understand the molecular properties, pathogenicity of wildfowl NDV and its relationship with poultry NDV, a NDV strain, NDV2, was isolated from wildfowl in 2011 and confirmed virulent using standard biological virulence tests. Nine pairs of specific primers were designed from sequence of NDV deposited in GenBank, and the complete genomic sequence of NDV2 strain was 15 192 nt（GenBank accession No. KF306265） and its genome comprised six ORFs, namely, encoded nucleocapsid protein（NP）, phosphoprotein（P）, matrix protein（M）, fusion protein（F）, haemagglutinin- neuraminidase（HN） and large protein（L）. The lengths of each gene were respectively 1 753, 1 451, 1 241, 1 792, 2 004 and 6 703 nt. NDV2 was classifid as Class II, genotype VII. Comparison of the homologies of each protein showed that NDV2 was highly homologous to several domestic genotype VII strains, such as Egret. GX. 11, Duck.WF.00 and Goose.GD450.11 in recent years, which was far higher than strains of other regions and other genotypes.These results suggested that NDV2 had high affinity to domestic strains. The JS 10 strain of Class I was the least homologous to NDV2. Sequence analysis of the F gene showed that 15 192 nt strains （including NDV2 strain） had a common motif for RRQKRF or KRQKRF which was consistent with highly pathogenic NDVs. F and HN genotyping showed consistency between NDV2 and the majority of the reference strains. However, the Indian. NDV4 strain from chicken, it belonged to genotype II for F gene, but for HN gene it belonged to genotype VII. Because NDV2 had start sequences identical to those of other reference strains, its start sequences were also highly conserved. The NP, P, M and L termination sequences of NDV2 were consistent with those of the reference strains but differed from those of the F and HN genes. NDV2 had termination sequences identical to those of L

关 键 词： 野禽 新城疫病毒 全基因序列 同源性比较 致病性

领　　域： [农业科学] [农业科学] [农业科学]