侵染万寿菊的黄瓜花叶病毒外壳蛋白基因的克隆与序列分析  

Cloning and Sequence Analysis of Coat Protein Genes of Cucumber mosaic virus Isolated from Tagetes erecta

作　　者：李晶晶[1] 邓召花[1] 曾静[1] 张金方[1] 张飞云[1] 

机构地区：[1]首都师范大学生命科学学院,北京100037

出　　处：《生物技术通报》2007年第6期92-96,100,共6页Biotechnology Bulletin

基　　金：国家自然科学基金面上项目(30571010);教育部科学技术研究重点项目(204006);北京市教委科技发展计划面上项目(KM200310028102)

摘　　要：应用ELISA的方法检测到感染了黄瓜花叶病毒(Cucumber mosaic virus,CMV)的万寿菊。根据已报道的CMV外壳蛋白(CP)基因的保守序列设计并合成引物,提取万寿菊叶片总RNA为模板,进行cDNA合成和PCR扩增,得到约875bp的片断,与预期片断大小相符。序列分析显示:该分离物CMV-WSJ CP基因全长657bp,编码218个氨基酸,其核苷酸和氨基酸序列与CMV亚组Ⅱ的分离物有很高的同源率,分别达到98.33%～99.24%和98.63%～100%;与CMV亚组I分离物的同源率分别仅为74.43%～76.26%和77.17%～78.99%。因此,从万寿菊叶片上检测出的黄瓜花叶病毒分离物CMV-WSJ应归属于亚组Ⅱ。Using ELISA method,we found that Tagetes erecta sample was infected by Cucumber mosaic virus. According to the sequences of coat protein genes of CMV formerly published,the primers were synthesized. A 875bp cDNA fragment was amplified by ILT-PCIL from the ILNA template of Tagetes erecta. The cDNA fragment was then cloned and sequenced. The analysis of sequences showed that the gene of CP had 657 nucleotides,encoding 218 amino acids. Compared with other isolations of CMV subgroup Ⅱ ,the sequence of nucleotides had about 98.33%-99.24% homology and the sequence of amino acids had 98.63%-100% homology;and compared with the subgroup Ⅰ isolations, each had only 74.43%-76.26% and 77.17%-78.99% homology. So sequence analysis revealed that the Cucumber mosaic virus isolated from Tagetes erect belonging to CMV subgroup Ⅱ.

关 键 词：万寿菊 黄瓜花叶病毒 外壳蛋白 CMV亚组Ⅱ 

分 类 号：S432.41[农业科学—植物病理学]