目录
摘 要 II
关键词 II
Abstract III
引言
引言 1
1 材料与方法 3
1．1 材料 3
1．2 方法 3
1．2．1 StHsfs 启动子区域EBOX和BRRE的分析 3
1．2．2 StBES12、AtBES1和AtBZR1氨基酸序列比对 3
1．2．3 StBES12、AtBES1和AtBZR1蛋白质二级结构、三级结构的预测及对比 4
1．2．4 马铃薯基因组提取及启动子序列克隆 4
1．2．5 双荧光素酶表达载体的构建 4
1．2．6 基因工程菌的构建和鉴定 4
2 结果与分析 5
2．1 StHsfs启动子区域内BES1结合基序EBOX和BRRE的分析 5
2．2 转录因子StBES12与转录因子AtBES1、AtBZR1氨基酸序列对比 6
2．3 转录因子StBES12与转录因子AtBES1、AtBZR1蛋白质二级结构预测及对比 6
2．4 转录因子StBES12与转录因子AtBES1、AtBZR1蛋白质三级结构预测及对比 7
2．5 双荧光素酶表达载体及基因工程菌的构建 8
3 讨论 9
3．1 同源基因与蛋白具有相似功能 9
3．2 后续实验计划 9
3．3 本实验的意义与应用前景 10
致谢 10
参考文献： 10
马铃薯转录因子StBES12与基因StHsfs启动子互作关系的初步探究
摘 要
BES1（BRI1EMSsuppressor 1）是一种广泛存在于植物中的转录因子,是植物响应油菜素甾醇（Brassinosteroid，BR）信号的响应元件，在调控植物生长发育和抗逆方面有着重要作用。基因Hsf（Heat shock factor）是植物抵抗热害等非生物胁迫的关键基因之一，它的表达受到诸多因素的调控，包括油菜素甾醇。在马铃薯中，外源施加BR缓解了叶片在高温下的萎焉状况，提高了其耐热性。但其分子机制却不清 *51今日免费论文网|www.jxszl.com +Q: *351916072* 
楚。早期工作已鉴定到27个StHsfs基因，其中部分受到高温的诱导。因此我们推测BR可能通过其下游转录因子StBES1直接促进StHsf的表达，从而提高马铃薯的耐热性。因此，本论文以StBES1与StHsfs启动子互作为研究内容,采用生物信息学的方法，对StHsfs启动子区域内EBOX和BRRE基序含量情况进行分析。同时我们也将StBES12和拟南芥中能与目标启动子结合的转录因子AtBES1 / AtBZR1进行蛋白质结构上的对比。之后我们在实验中构建双荧光素酶表达载体并将其转化进基因工程菌。这些结果为后期蛋白质DNA互作实验提供思路与准备，从而为解析油菜素甾醇通过StBES1直接调控StHsfs表达的分子机制打下基础。
PRELIMINARY STUDY ABOUT INTERACTION BETWEEN TRANSCRIPTION FACTOR StBES12 AND GENE StHsfs PROMOTER IN Solanum tuberosum L.
ABSTRACT
BES1 (BRI1EMSsuppressor 1) is an extensive transcription factor in plants. It is an important responsive element of brassinosteroid (BR) signal, which plays an important role in stress resistance, growth and development of plants. Hsf (Heat shock factor) is a key gene which can resist heat stress and the other abiotic stress. Its expression is regulated by many factors, including BRs. Exogenous BR can alleviate the wilting condition of potato leaves at a high temperature and improve its heat resistance. However, the molecular mechanism is not clear. Twentyseven StHsfs members were identified in earlier work, and some of them are responsive to heat stress. Therefore, we deduced that BR can induce the expression of StHsfs through StBES1, further to improve the heat resistance of potato. Based on this hypothesis, this study takes StBES1 and StHsfs promoters interaction as research content. We used bioinformatics to analyze the contents of EBOX and BRRE motifs in StHsfs promoters, while we also analyzed the protein structure of StBES12 compared with the known AtBES1/AtBZR1 which can binds to the target promoters in Arabiodpsis. Then we constructed the plasmid with luciferase expression system and transformed it into genetically engineered bacteria. These results provide ideas and preparation for the proteinDNA interaction experiment, thus laying the foundation for the analysis of the molecular mechanism of Brassinosteroids regulating StHsfs expression through StBES1.

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