基因型鹅副粘病毒NA-1株全基因组的克隆及特性分析  

Cloning complete genome and characteristic analysis of genotype Ⅶ goose paramyxovirus strain NA-1

作　　者：徐明[1] 丁壮[1] 毕玉海[1] 李志杰[1] 常爽[1] 黄海楠[1] 宋子运[1] 尹仁福[1] 杜眉[1] 

机构地区：[1]吉林大学畜牧兽医学院,吉林长春130062

出　　处：《中国兽医学报》2006年第6期610-613,618,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(30571375)

摘　　要：将本实验室分离保存的NA-1株鹅副粘病毒（GPMV）经SPF鸡胚增殖，收集鸡胚尿囊液进行病毒纯化，提取病毒基因组RNA。参考GenBank已收录的GPMV ZJ1株基因组序列，设计了8对特异性引物，RT—PCR法分别特异性的扩增出病毒各基因片段，并将各目的基因片段回收纯化．克隆PGEMT载体．转化大肠杆菌DH5α，小提质粒选取阳性克隆酶切鉴定并进行序列测定．对测定结果进行拼接得到全长cDNA序列（GenBank登录号：DQ659677）。NA-1株核苷酸比新城疫病毒（NDV）核苷酸多6个碱基，位于1644～1645nt之间，符合NDV核苷酸“六碱基原则”。序列分析结果表明，GPMVNA-1株与各地代表株核苷酸同源性81．2％～91．3％。同时根据各毒株F基因开放阅读框前389个核苷酸序列绘制出系统发育进化树，结果表明GPMNNA-1株与SFO2和QY97病毒株同属于基因Ⅶ型，不同于基因Ⅸ型NDV的国家标准株F48E9和基因Ⅱ型的LaSota传统弱毒株。Genotype Ⅶ goose paramyxovirus（GPMV）strain NA-1 was separated and conserved by this laboratory, Virus was multiplied by SPF embryonated eggs to collect allantoie fluid in order to obtain the purified virus, and then to extract the RNA genome of virus, Based on the complete genome sequence of goose paramyxovlrus strain ZJ1 logged in GenBank and then eight pairs specificity primers were designed to amplified every gene fragment by the method of RT-PCR, Meanwhile, the target gene fragments were reclaimed and purified to insert into PGEM-T vector to converted into the E, coli DH5a, Slight plasmid was extracted to select the masculine products for identity by enzyme digestion, The complete genome after sequencing and splicing was logged in the GenBank（Number：DQ659677）, The base number of nucleotide from GPMV is six more than the base number of nucleotide from NDV which accords with the ＂the six base principle＂. Sequence analysis results indicated that the homology of complete genome nucleotide sequence from strain NA-1 was between 81.2% and 91.3% as compared with representative strains from every locals. Meanwhile, on the basis of the nucleotide sequence of F gene from each NDV strains in a 389 bp region, the phylogenetic tree was drawn, that indicated goose paramyxovirus virulent strain NA-1 belonged to genotype Ⅶ which was the same with virulent strain SFO2 and strain QY97, but different from NDV virulent strain F48E9 as the government standard strain in China which belongs to genotype Ⅸ and the traditional low virulent strain Lasota which belongs to genotype Ⅱ .

关 键 词：基因Ⅶ型鹅副粘病毒 新城疫病毒 全基因组 

分 类 号：S852.65[农业科学—基础兽医学]