摘要
以新城疫病毒 ( NDV)长春株接种 9日龄 SPF鸡胚增殖后 ,进行了蚀斑纯化 (三代 )鉴定 ,差数、蔗糖密度梯度离心提纯和生物学特性鉴定。结果表明 ,该病毒的鸡胚平均致死时间 ( MDT)大于 1 6 8h,雏鸡脑内致死性 ( ICPI)为 0 .2 5,雏鸡静脉致死性 ( IVPI)为 0 ,血凝解脱时间超过 1 2 0 h,血凝素热稳定性 56℃ 5min仍具有血凝活性。系统发育进化树分析表明 ,NDV长春株与 La Sota株亲缘关系最近 ,为弱毒株。参考多株 NDVHN基因序列设计了 1对特异性引物 ,应用 RT-PCR一次性扩增出 NDV长春株的全长 HN基因。将该 HN基因插入 p KS( -)后 ,进行了序列测定。序列分析表明 ,该 HN基因核苷酸长度为 1 731 bp,编码 577个氨基酸 ,序列中有 5个糖基化位点 ,1 2个半胱氨酸残基 ,与国外发表的 NDV毒株序列相符 ,核苷酸同源性为88.1 %～ 98.8% ,推导的氨基酸同源性为 91 .1 %～ 98.8%。
The NDV strain Changchun was grown in 9 days SPF chicken embryos. The virus was purified and the genomic RNA was extracted. The study of biological character revealed that the mean death time (MDT) of chicken embryos was more 168 h and introcerebral pathogenicity index(ICPI) in 1 day old chicken was 0.25 and intravenous pathogenicity index(IVPI)in 6 week old chicken was 0. Phylogenetic tree showed the more relationship between the NDV strain Changchun and La Sota. The results showed the NDV strain Changchun was avirulent. A pair of special oligonucleotide primers were generated by HN gene sequence of NDV strain Texas GB and Italien and whole sequence of HN gene of NDV strain Changchun was amplified by RT PCR. The HN gene was sequenced after being directly inserted in plasmid pKS( ) to be a nucleotide sequence of 1 731 bp encoding a protein of 577 amino acids. In the amino acid sequence,there are 5 glycosylation sites and 12 cysteine residues. Compared with the published NDV strains,the homology of the nucleotide sequence is 88.1% 98.8% and the homology of the deduced amino acid sequence is 91.1% 98.8%.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2000年第6期539-543,共5页
Chinese Journal of Veterinary Science
基金
国家自然科学基金重大项目! ( 398932 90 )
国家"86 3"计划资助项目 !( 10 1-0 5 -0 3-1)
