鹅副粘病毒NA-1分离株HN蛋白基因的克隆与序列分析  

Cloning and sequenceing of HN gene of avian paramyxovirus strain NA-1 isolated from goose

作　　者：王学理[1,2] 丁壮[1] 左玉柱[1] 向华[1] 常爽[1] 黄海楠[1] 宣华[1] 

机构地区：[1]解放军军需大学兽医医院 [2]内蒙古民族大学动物科技学院,内蒙古通辽028000

出　　处：《中国预防兽医学报》2005年第1期18-21,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：吉林省科委发展计划项目 (2 0 0 0 0 0 2 0 0 )

摘　　要：鹅副粘病毒分离株NA_1经 11日龄鸡胚增殖后纯化。提取病毒的基因组RNA ,采用RT_PCR扩增出与预期设计的 1 7kb大小相符合的特异条带。将扩增产物提纯后克隆入pMD 18_T载体 ,经纯化、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 ,并对阳性克隆进行测序。测序后拼接得出HN基因的全序列长度为 174 0bp ,该基因的ORF总长为 1716bp ,编码 5 71个氨基酸。与GenBank下载的 15株参考毒株比较HN基因编码区全核苷酸序列 ,发现所测NA_1毒株与参考毒株YG97核苷酸序列同源性为 97 3% ,与F48E9同源性为 84 5 % ,与LaSota为 82 2 %。Avian paramyxovirus strain NA-1 isolated from goose was propagated in 11-day-old chicken embryos.The genomic RNA of NA-1 were extracted.The HN gene of the virus has been successfully amplified by RT-PCR,and further cloned into pMD 18-T vector,and a positive recombinant plasmid was obtained by restriction enzyme analysis. The complete sequence of HN gene was obtained finally and the nucleotide sequence of the HN gene was 1 740 bp.The sequence analysis demonstrated that 15 reference strains loaded down from GenBank,the result of sequence analysis indicated that NA-1 strain exhibited 97.3?% homology with reference strain YG 97,84.5?% homology with strain F-(48)E-9 and 82.2?% homology with strain La Sota. The results indicated the mutation of the HN gene between the goose's paramyxovirus and NDV varied to a great extent.

关 键 词：鹅副粘病毒 NA-1分离株 HN蛋白 基因克隆 序列分析 同源性 

分 类 号：S852.659.5[农业科学—基础兽医学]