犬2型腺病毒六邻体分子特性分析.PDF

犬2 型腺病毒六邻体分子特性分析1 王芳 李昌文 葛俊伟 张洪英 李建伟 陈洪岩 曲连东* 中国农业科学院哈尔滨兽医研究所黑龙江 哈尔滨150001 摘要 目的 分析犬2 型腺病毒六邻体分子特性 方法 参考GenBank 发表的犬2 型腺病毒 CAV-2 基因序列 设计并合成1 对引物 采用PCR 方法对CAV-2 疫苗株HSY 的主要衣壳蛋白六邻体 Hexon 基因进行扩增 将扩增产物与pMD 18-T 载体连接 转化 筛选出阳性克隆进行测序 结果 CAV-2 疫苗株HSY 六邻体的核苷酸序列全长27 18bp 编 码906 个氨基酸 与TorontoA26/6 1 株相比 核苷酸同源性为99.85% 推导氨基酸序列同源 性为99.7% 有4 个核苷酸发生变异 引起2 个氨基酸的改变 但相应区域的氨基酸疏水性 并未发生明显改变 和CAV-1 六邻体基因序列比较 核苷酸同源性为88 左右 氨基酸同 源性为96 左右 和其他哺乳动物腺病毒株六邻体基因比较 核苷酸同源性在50.2 75.5 之间 氨基酸同源性在 49.9 75.3 之间 结论 克隆获得六邻体全基因序列 进化 树和同源性分析表明 CAV-1 和CAV-2 同源性较高 亲缘关系最近 和牛腺病毒最远 关键词 犬2 型腺病毒 六邻体 克隆 序列分析 中国分类号 文献标志码 文章编号 Analyzing of molecular character hexon gene of canine adenovirus type 2 WangFang,Changwen Li,Junwei Ge,Hongying Zhang,Jianwei Li, ,Hongyan Chen Liandong Qu * Harbin Veterinary Research Institute of CAAS Harbin 150001 ) Abstract Objective To analysis character of CAV-2 hexon gene. Methods According to the reported complete nucleotide sequence of canine adenovirus in GenBank, a pairs of primers were designed and synthesized. By the method of PCR,the hexon gene of CAV-2 HSY strain was amplified.In order to further identify the credibility of the hexon gene,the products of PCR was cloned into pMD 18-T vector and sequenced. Result Full length of hexon gene was 2718nt and encoded 906 amino acids.Compared the hexon gene of CAV-2 HSY strain with TorontoA26/6 1,it showed that they share 99.85% identity in nucleotide sequence and 99.7% identity in deduced amino acid sequence separatedly.There are four nucleotides changed that changed two amino acids, but corresponding amino acids didn’t chang