论文部分内容阅读
用CTAB法从感染草莓镶脉病毒(strawberry vein banding virus,SVBV)的草莓叶片中提取总DNA,设计特异性引物扩增SVBV ORF Ⅵ基因的全长片段,进行克隆并测序.序列分析表明SVBV ORF Ⅵ基因完整序列全长1 557bp,编码518个氨基酸.将其与SVBV美国分离物以及花椰菜花叶病毒属其他成员的ORF Ⅵ基因相比较,结果表明SVBV中国分离物ORF Ⅵ基因与SVBV美国分离物ORF Ⅵ基因核苷酸序列相似性最高,达84.5%;而与花椰菜花叶病毒属其他成员的ORF Ⅵ基因序列相似性均较低,仅为23.7%～27.9%.构建SVBV及其同属其他成员ORF Ⅵ基因的系统关系树,结果显示SVBV中国分离物与SVBV美国分离物单独形成1个分支,说明来源于草莓的2个SVBV亲缘关系最近,而与其同属其他成员的亲缘关系均较远.生物信息学分析表明SVBVORF Ⅵ基因表达的P6蛋白序列含有多种蛋白激酶磷酸化位点,其二级结构N端是1个α-螺旋富集区.该研究将为进一步研究P6蛋白的功能提供理论依据.
Total DNA was extracted from strawberry leaves infected with strawberry vein banding virus (SVBV) by CTAB method, and specific primers were designed to amplify the full-length fragment of SVBV ORF Ⅵ gene, then cloned and sequenced.The sequence analysis showed that SVBV ORF The complete nucleotide sequence of Ⅵ gene was 1 557 bp in length and encoded a polypeptide of 518 amino acids.Compared with other isolates of SVBV and other members of the genus Cauliflower mosaic virus, the ORF Ⅵ gene of SVBV isolate showed that ORF Ⅵ gene of SVBV isolates from China was associated with SVBV isolates The nucleotide sequence of ORF Ⅵ gene had the highest similarity (84.5%), but the ORF Ⅵ gene sequence similarity with other members of Cauliflower mosaic virus was low (23.7% -27.9%). The construction of SVBV and its other members ORF Ⅵ gene system showed that SVBV isolates from China and SVBV isolates formed a single branch, which indicated that the two SVBVs derived from strawberry had the closest genetic relationship, but were distantly related to other members of the same family. The informatics analysis showed that the P6 protein sequence of SVBVORF Ⅵ gene contains many protein kinase phosphorylation sites and the N-terminal secondary structure is an α-helix-rich region. Further studies of protein P6 provide a theoretical basis.