鹅副粘病毒YG_(97)分离株HN蛋白基因的克隆与序列分析

Cloning and Sequencing of HN Gene of Goose's Paramyxovirus

鹅副粘病毒分离株YG97经 10日龄鸡胚增殖后纯化 ,提取病毒的基因组RNA ,采用RT_PCR一次性扩增出与预期设计的 1.9kb大小相符合的特异性条带。将扩增产物提纯后克隆入pGEMR_T载体 ,经转化、筛选及酶切鉴定后 ,初步获得了含鹅副粘病毒HN基因的阳性克隆 ,进一步进行了序列测定。序列分析表明所扩增的病毒的HN基因片段的长度为 1981bp ,共编码5 71个氨基酸。同源性分析表明 ,YG97与LaSota毒株核苷酸同源性为 79% ,氨基酸的同源性为 87%。与台湾 1995年分离株Taiwan/ 95核苷酸和氨基酸的同源性分别为 93%、96 % ,说明YG97与Taiwan/ 95亲缘关系较近 ,具有较高的相似性。与国内外部分发表的其它NDV毒株的核苷酸同源性在 80 %～ 84%之间 ,氨基酸的同源性在 87%～ 91%之间。

The genomic RNA of YG 97 were extracted.The hameagglutinin-neuraminidase (HN) gene of YG 97 has been successfully amplified RT-PCR and coloned in to pGEM-T vector.The sequence analysis showed that the nucleotide sequence of the HN gene was 1981 bp and encoding a protein of 571 amino acids.Sequence comparison between YG 97and the Taiwan/95 strains,the homology of the nucleotide was 93%,the homology of the amino acid was 96%,which indicated they probably had a common origin.The homology of the nucleotide compared with LaSota was 79%,the homology of the amino acid was 87%.The results indicated the mutation on the HN gene had varied largely,compared with classic NDH.