【摘要】 以犬瘟热病毒水貂分离株的RNA为模板,对分离株的H基因进行RT-PCR扩增,扩增产物纯化后与pMD-18T SimpleVector连接,成功地构建了克隆质粒pMD-18T-CDVH。序列分析表明,分离株H基因由1596 bp组成,编码532个氨基酸,含有5个潜在的N-联糖基化位点,12个半胱氨酸残基。与已发表的24株CDV毒株进行同源性比较,核苷酸序列同源性在90.7%～98.6%之间;推导的氨基酸序列同源性在89.9%～97.0%之间。与Onderstepoort、Convac、Hamatsu、A75/17、TN株相比,根据推导的氨基酸序列比较分析表明,N-联糖基化位点的数目与位置均存在差别。聚类分析结果表明分离株与Snyder Hill毒株的亲缘关系最近,二者与疫苗株Onderstepoort和Convac具有较高的同源性,组成一组,而与标准野毒强毒株Hamatsu的亲缘关系较远。更多还原

【Abstract】 Hemagglutinin(H) gene of canine distemper virus isolated from mink was amplified with RT-PCR using total RNA from the strain as template,and the RT-PCR products were purified.The purified products were linked with pMD-18T Simple Vector,creating the recombined cloning plasmid pMD-18T-CDVH.Sequencing results indicated that H gene section of the isolated strain was of a 1 596 bp fragment encoding 532 amino acids,containing 5 potential N-glycosylation locations and 12 cysteine residues.Compared with 24 published CDV strains,the nucleotide sequence homology was among 90.7%～98.6%.The Derived amino acid homology was among 89.9%～97.0%.Compared with Onderstepoort,Convac,Hamatsu,A75/17 and TN strains,there were differences in the number and locations of N-glycosylation based on the derived amino acid sequences.Clustering analysis results showed that the isolated strain was closely related to the Snyder Hill strain,forming one group,and sequence homologies of both strains with vaccine strains Onderstepoort and Convac were high.However,the genetic relationship between them and standard virulent strain Hamatsu was far away.更多还原