鸡传染性支气管炎病毒嗜腺胃毒株(ZJ_(971))核衣壳蛋白基因的克隆  

Cloning of Nucleocapsid Protein Gene of Proventriculopathic Infectious Bronchitis Virus ZJ_(971) Isolate

作　　者：丁红梅[1] 周继勇[1] 沈行燕[1] 吴建祥[1] 程丽琴[1] 于涟[1] 

机构地区：[1]浙江大学动物预防医学研究所,浙江杭州310029

出　　处：《农业生物技术学报》2001年第3期243-246,共4页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(39970030);浙江省自然科学基金(399411);浙江省重点科技项目(991102030)资助

摘　　要：从以鸡腺胃肿大为特征的病例中鉴定了一株新的禽传染性支气管炎病毒变异株，命名为传染性支气管炎病毒嗜腺胃毒株，ＺＪ９７１毒株。应用ＲＴ－ＰＣＲ方法对传染性支气管炎病毒嗜腺胃毒株（ＺＪ９７１）的核衣壳蛋白基因（Ｎ基因）进行扩增，将扩增得到的Ｎ基因ｃＤＮＡ克隆入ｐＢｌｕｅｓｃｒｉｐｔＳＫ（＋）质粒载体中，经鉴定后测序。用ＰＣＧＥＮＥ软件对测得的序列进行分析。结果显示，ＺＪ９７１毒株Ｎ基因ＯＲＦ全长为１　２３０　ｎｔ，编码一条４０９个氨基酸组成的多肽，与Ｂｅａｕｄｅｔｔｅ株、Ｍ４１株、Ｇｒａｙ株相比较，其核苷酸同源性分别为９０．９８％、９０．６５％、９２．２８％，氨基酸同源性分别为９２．１８％、９２．４２％和９２．９１％。ＺＪ９７１毒株Ｎ基因核苷酸与Ｂｅａｕｄｅｔｔｅ株、Ｍ４１株、Ｇｒａｙ株的不同之处是其５４５～５９５位核苷酸的突变率分别为２６％、２６％和１６％。ＺＪ９７１毒株Ｎ基因的变异特征不同于Ｓ１基因。A novel proventriculopathogic variant ZJ971 isolate of infectious bronchitis virus(IBV)had been identified from the sick chickens with the enlarged proventriculi.The nucleocapsid protein(N)gene cDNA segement of 1.23 kb was amplified by RT-PCR with the viral RNA of the ZJ971 isolate.The cDNA was cloned into pBluescript SK(+)vector.Nucleotide sequence of the open reading fragment(ORF)of N gene of ZJ971 isolate was 1 230 nt,coding a polypeptide of 409 amino acids.Comparison of sequence of N gene of ZJ971 isolate with strain Beaudette,M41 and Gray showed that the nucleotide homologies were 90.98%,90.65% and 92.28%,and the amino acids homologies were 92.18%,92.42% and 92.91% respectively.The remarkable characteristic of the N gene of ZJ971 isolate was that its mutation rates were as high as 26%,26% and 16% respectively between 545～595 regions.Variant characteristic of N gene of IBV-ZJ971 isolate was different from its S1 gene.

关 键 词：传染性支气管炎病毒 核衣壳蛋白 基因克隆 序列分析 鸡 嗜腺胃毒株 

分 类 号：S852.65[农业科学—基础兽医学] S858.315.3[农业科学—兽医学]