水稻oscpk9基因启动子的克隆及功能分析-cloning and functional analysis of promoter of rice os cpk 9 gene.docxVIP

华 华 中 科 技 大 学 硕 士 学 位 论 文 II万方数据 II 万方数据 动子活性，且表达在不同的组织中存在着差异性。 （3）采用农杆菌介导的瞬时表达技术分析表明，植物激素脱落酸（ABA）和渗 透物质 PEG-6000 对 POsCPK9 驱动的 GUS 基因的表达有诱导作用，经过 3 μmol/L ABA 和 20% PEG6000 处理的烟草叶片，其 GUS 活性较未诱导叶片均有不同程度的 提高。 （4）利用构建成功的 POsCPK9 ::GUS 真核表达载体遗传转化水稻，得到 20 株 转基因植株。经过 PCR 分子鉴定，发现 GUS 标记基因已经整合进部分转基因植株中， 初步筛选到了 4 株 T0 代转基因阳性植株，为后续深入研究 OsCPK9 表达调控模式的 实验奠定了基础。 关键词：OsCPK9；启动子；GUS；瞬时表达；水稻；烟草；遗传转化; III万方数据 III 万方数据 Abstract Calcium, as one of the important second messengers, plays an important role in plant growth and developmental processes and in response to environmental stresses. To date, several classes of calcium-sensing proteins have been characterized in plants. Among them, calcium-dependent protein kinases (CDPK) attracts special attention because it acts not only as the Ca2+ sensor, but also the effector, and is widely distributed in plants. In recent years, CDPKs have been shown to be involved in the plant development, hormone responses, metabolism, environmental stresses, defense responses and other processes. It was found OsCPK9 transcript can be induced by drought, high salt stresses and rice blast infection, indicating that OsCPK9 may play important roles in response to rice bio- and abiotic stresses. Promoter, as cis-acting elements, is located upstream of the transcription initiation site. It can be recognized and binding with RNA polymerase, there by initiating the downstream gene expression. It determines temporal and spatial specificities of gene expression in plants, and therefore is called molecular switch”. In order to investigate regulation mechanism of the OsCPK9 gene, we cloned and sequenced about 2 kb upstream sequence of rice OsCPK9 gene transcription initiation site. Cis-element analyses were performed with PLACE database. POsCPK9::GUS expression vector was successfully constructed for studying OsCPK9 gene expression characteristics. The main results are as follows: The 2114-bp long upstream sequence of OsCPK9 gene transcription initiation