两株大蒜病毒外壳蛋白基因的克隆及其在大肠杆菌中的表达  

CLONING AND EXPRESSION OF TWO GARLIC VIRUS COAT PROTEIN GENES

作　　者：马筠[1] 杨恭[2] 徐绍华[2] 魏军亚[2] 邱并生[2] 

机构地区：[1]天津师范大学化学与生命科学院,天津300074 [2]中国科学院微生物研究所,北京100080

出　　处：《微生物学报》2001年第4期415-420,共6页Acta Microbiologica Sinica

基　　金：天津市二十一世纪青年基金赞助! (963 70 2 81 1 )&&

摘　　要：利用建立cDNA库的方法 ,通过RT PCR分别扩增和克隆了感染我国天津地区大蒜的大蒜花叶病毒 (GMVc)和大蒜潜隐病毒 (GLVc)的外壳蛋白 (CP)基因 ,并对其分别在原核细胞中进行了诱导表达和表达产物的分析。克隆基因的序列测定与分析表明 ,GMVc的CP基因全长 867个核苷酸 ,编码 2 89个氨基酸 ,与报道的一株GMV的核苷酸和氨基酸的同源性分别为 88 5%和 97 2 % ,属于马铃薯Y病毒属 (Potyvirus)的成员 ;GLVc的CP基因全长 885个核苷酸 ,编码 2 94个氨基酸 ,与报道的一株GLV的核苷酸和氨基酸同源率分别为 73 6%和 90 9% ,属于香石竹潜隐病毒属 (Carlavirus)的成员 ;克隆基因的表达产物经SDS PAGE分析表明 ,GM Vc和GLVc的外壳蛋白分别约 32kD和 34kD ,与预测的结果一致。本实验结果对感染我国大蒜的病毒进行分子水平的确切诊断、分类和调查奠定了基础 ,将对大蒜病毒病的防治与脱毒大蒜的生产具有重要的意义。The coat protein(CP)genes of garlic mosaic virus(GMVc)and garlic latent virus(GLVc)isolated from garlic( Allium) plants in Tianjin,China,were amplified from an established cDNA library by PCR method and subsequently expressed in E.coli. using the pET\|30a expression system.The determined sequences of GMVc and GLVc CP genes show that the complete GMVc CP gene has 867 nucleotides encoding 289 amino acids.It has 88.5% and 97.2% homology,at the levels of nucleotide and amino acid,respectively,to a reported GMV,indicating that it belongs to Potyvirus. The complete GLVc CP gene has 885 nucleotides coding for 294 amino acids.It has 73.6% and 90.9% homologous percents,in nucleotide and amino acid,respectively,compared to a previously reported GLV,suggesting that it is a member of Carlavirus .The expressed products presented in inclusion body and were analyzed by SDS\|PAGE.The molecular weights of GMVc and GLVc CPs appear in 32kD and 34 kD size,respectively,which are consistent with the deduced sizes of these two CPs.These data will be virtually significant to the further investigation of viruses infecting parlic plant,the control of garlic virus diseases and the production of virus\|freed garlic plants.

关 键 词：大蒜花叶病毒 大蒜潜隐病毒 外壳蛋白基因 克隆 表达 

分 类 号：Q786[生物学—分子生物学]