鹅副粘病毒NA-1分离株HN蛋白基因的克隆与序列分析

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1 740bp,该基因的ORF总长为1 716 bp,编码571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La Sota为82.2%.同源性分析表明NA-1相对于NDV在HN基因上发生了较大的变异.

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

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.