滇黑两省黄瓜花叶病毒南瓜分离物外壳蛋白基因序列分析

 利用免疫捕捉多聚酶链式反应( Immuno-Capture PCR, IC-PCR) 方法从南瓜的两个黄瓜花叶病毒(Cucumber mosaic virus, CMV) 云南分离物(CMV-YN) 和黑龙江分离物(CMV-HLJ ) 中扩增获得约700 bp的外壳蛋白(Coat protein, CP) 基因片段, 并克隆到pGEM-T载体中。核苷酸序列测定表明, CMV-HLJ和CMV-YN两个分离物CP基因长为657 bp, 编码219个氨基酸, 两个分离物的核苷酸和氨基酸同源性分别达到96.8%和97.4%。和国外CMV亚组Ⅰ 6个分离物核苷酸序列同源性在90%以上, 氨基酸同源性在97%以上, 和亚组Ⅱ 6个分离物的同源性分别为66.4%和73.4%。和国内亚组Ⅰ10个分离物的核苷酸同源性都在89.5%以上, 氨基酸同源性在93%以上, 而和亚组Ⅱ两个分离物的同源性分别为68.3%和73.5%。序列分析结果表明CMV-HLJ和CMV-YN两个分离物属于CMV亚组Ⅰ。

Sequence Analysis of CP Genes of CMV from Pumpkin in Yunan and Heilongjiang Provinces

The CP genes were amplified by IC-PCR from CMV-HLJ and CMV-YN isolated from pumpkin in Heilongjiang and Yunan Provinces and cloned to pGEM-T vector. Analysis of nucleotide sequence showed that the length of CP genes of CMV-HLJ and CMV-YN was 657 bp, encoding 219 amino acid. The homologyof nucleotide and amino acid sequences of CP gene between these two isolates was 96.8% and 97.4% respectively. The homology of nucleotide and amino acid sequences of CP gene of these two isolates was higher than90% and 97% respectively compared with six external isolates of CMV subgroupⅠ, and 66.4% and 73.4% respectively compared with another six external isolates of CMV subgroup Ⅱ, and higher than 89.5% and 93% respectively compared with ten Chinese isolates of CMV subgroup Ⅰ, and 68.3% and 73.5% respectively compared with two Chinese isolates of CMV subgroup Ⅱ. All these data showed that CMV-HLJ and CMV-YN belonged to CMV subgroupⅠ.