国产IBV疫苗株核衣壳蛋白基因的亲缘关系  

Genetic Relationship of Nucleocapsid Gene of Domestic IBV Vaccine Strains

作　　者：曹伟胜[1] 廖明[1] 任涛[1] 罗开健[1] 辛朝安[1] 

机构地区：[1]华南农业大学兽医学院,广东广州510642

出　　处：《中国兽医学报》2003年第2期119-123,共5页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金重大项目 (3 9893 2 90 -2 )

摘　　要：利用自行设计的引物 Cx和 Cs,通过 RT- PCR方法分别扩增出鸡传染性支气管炎病毒 (IBV) D4 1株、H12 0 GD株、H12 0 SH株、H5 2 GD株等 4个国产疫苗株和标准强毒 M4 1- E4株完整的 N基因 c DNA,然后将其分别克隆到p GEM T- Easy或 p MD18- T载体中并测序。测序结果表明 ,这 5个 IBV毒株的 N基因均为 12 30 bp,分别编码由 4 0 9个氨基酸残基组成的多肽。同源性分析发现 ,被检的 5株 IBV毒株可分 2组 ,其中 D4 1株、H12 0 GD株和 H12 0 SH株为一组 ,它们的核苷酸和推导氨基酸序列同源性为 99.7%～ 99.8%和 99.0 %～ 99.5 % ,而 H5 2 GD株与 M4 1- E4株构成另一组 ,其核苷酸和推导氨基酸序列同源性为 99.7%和 99.5 % ;而 2组之间的最大同源性仅为 91.0 %和 93.2 %。在系统发生进化树上 ,这 2组分别位于不同的分支簇上。值得注意的是 ,国内的 H5 2 GD株与国外报道的 H5 2株不在同一分支簇上 ,相反却与国内强毒 M4 1- E4株以及国外报道的 M4 1株在同一分支簇上。这一结果表明 ,国内的 H5 2 GD疫苗株与国外报道的 H5 2疫苗株不同 ,它们在亲缘关系上更靠近 M4 1- E4株和 M4 1株。The nucleocapsid protein gene(N gene) of IBV strain D41,H120GD,H120SH,H52GD and M41-E4 were amplified by RT-PCR with a pair of specific oligonucleotide primers(Cx/Cs) and cloned into plasmid pGEM-T Easy and pMD 18-T respectively.The inserts were sequenced and the results revealed that the complete N genes of strains D41,H120GD,H120SH,H52GD and M41-E4 were all composed of 1 230 bp,coding for 409 amino acids respectively.Sequence analysis revealed that the five IBV strains could be divided into two groups,i.e.,strains D41,H120GD and H120SH formed one group and there was 99.7%-99.8% nucleotide sequence homology and 99.0%-99.5% deduced amino acid sequence homology between strains D41,H120GD and H120SH;while strains H52GD and M41-E4 constituted another group and they had 99.7% nucleotide sequence homology and 99.5% deduced amino acid sequence homology each other.But the highest homology between the two groups was only 91.0% and 93.2%.The two groups fell into two different branches of a phylogenetic tree.It was noteworthy that domestic vaccine strain H52GD was not in the same branch with foreign H52 strain,but with domestic virulent M41-E4 strain and foreign M41 strain.The study suggested domestic H52 strain(H52GD) was different from foreign H52 strain because the former was more genetically related to domestic virulent M41-E4 strain and foreign M41 strain.

关 键 词：核衣壳蛋白基因 亲缘关系 鸡 传染性支气管炎病毒 疫苗株 N基因 序列分析 

分 类 号：S852.659.5[农业科学—基础兽医学]