| 摘要: |
| 【目的】对草莓镶脉病毒（SVBV）启动子进行序列测定和分析，为进一步研究该启动子稳定表达活性、表达类型及驱动外源基因稳定表达提供理论依据。【方法】用CTAB法从感染SVBV的草莓叶片中提取总DNA，设计特异性引物扩增SVBV启动子，克隆并测序。将SVBV启动子与花椰菜花叶病毒属其他成员的启动子核苷酸序列进行比较，并构建其系统关系树，再用PlantCARE软件分析SVBV启动子序列中的各个作用元件。【结果】获得全长为1017 bp的SVBV 启动子。序列比列表明，本研究构建的中国SVBV启动子与美国SVBV启动子序列相似性最高，达77.29%。从构建的系统关系树可以看出，中国SVBV启动子与美国SVBV启动子单独聚成一个亚分支，说明来源于草莓的2个SVBV启动子亲缘关系最近。另外，SVBV启动子和花椰菜花叶病毒（CaMV）启动子亲缘关系虽然相对较近，但二者序列相似性较低，说明SVBV启动子与CaMV 35 S启动子的结构差异较大。进一步分析表明，SVBV启动子除了具有一般植物启动子的典型作用元件TATA-box和CAAT-box外，还具有一些与组织特异性表达或诱导表达相关的调节因子。【结论】SVBV启动子具有多种转录顺式作用元件，可能是一个在双子叶植物中具有较强驱动活性的组成型启动子。 |
| 关键词: 草莓镶脉病毒 启动子 克隆 序列分析 |
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| 基金项目:国家自然科学基金项目(30740033)；安徽省科技厅自然科学基金项目（11040606M68）；安徽省教育厅自然科学基金重点项目 (KJ2009A105，KJ2008A133) |
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| Cloning and sequence analysis of promoter of Strawberry vein banding virus（SVBV） |
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| Abstract: |
| 【Objective】The study was to sequence and analyze the promoter of Strawberry vein banding virus（SVBV） to provide theoretical basis for further researching steady expression activities, expression type and stability of driving exogenous gene expression of SVBV promoter. 【Method】The total DNA was extracted from strawberry leaves infected with SVBV by CTAB method.Specific primer pair was designed to amplify SVBV promoter,and then the promoter was cloned and sequenced.The sequence of SVBV promoter was compared with that of the promoter of other members of Caulimovirus,and a phylogenetic tree was constructed based on alignment of the nucleotide sequences of promoters of SVBV and other members of Caulimovirus.Functional elements of SVBV promoter was analyzed by software PlantCARE.【Result】An 1017 bp full-length sequence of SVBV promoter was acquired.Sequence comparison showed that Chinese SVBV promoter shared the highest sequence similarity (77.29％) with that of the American SVBV.Phylogenetic tree indicated that the Chinese SVBV promoter and the American SVBV promoter clustered into a separate subgroup.It illustrated that the two SVBV promoters derived from strawberry had closest relationship.Although the two SVBV promoters had relatively close relationship with CaMV promoters,they shared relatively lower similarity.It means that the structure of SVBV promoter greatly differs from the 35S promoter of CaMV.Further analysis showed that SVBV promoter contained some functional elements related to tissue specific expression and inducible expression besides typical functional elements TATA-box and CAAT-box of general plant promoter.【Conclusion】SVBV promoter contains various cis acting elements for transcription,and probably is a constitutive promoter having stronger expression activities in dicotyledons. |
| Key words: Strawberry vein banding virus promoter cloning sequence analysis |
