玉米抗冷相关基因ZmRBP1的克隆及表达载体构建.docxVIP

PAGE \* MERGEFORMAT PAGE \* MERGEFORMAT III 中文摘要 玉米的储备量位居我国粮食作物前列，其产量在我国粮食安全中起着关键作用。由于人们对玉米的需求量不断增加，常规的育种方法培育出的玉米种质无法满足目前的需求。因此，利用现代生物技术改良玉米的性状、创制新材料，提高玉米在严峻条件下的耐受能力成为最为目前的迫切需要。由于东北地区纬度略高，相对寒冷，易受到冷空气影响，对玉米及其他农作物产量构成威胁，因此开发及研究玉米抗冷相关基因成为必然趋势。 玉米抗冷相关基因ZmRBP1(Ran binding protein 1—Rbp1)所编码蛋白为Ran结合蛋白，定位于细胞质中，核苷酸全长为1467bp，能够编码488个氨基酸。在抗逆、激素信号传导以及植物生长发育过程中发挥重要作用。 本研究以玉米抗冷自交系W9816为材料，设计特异引物，利用RT-PCR技术，克隆得到RBP1基因。通过对RBP1氨基酸序列的生物信息学分析，与其他植物氨基酸的同源性达到80%以上，表明该序列为正确的RBP1基因序列。本实验进一步通过从克隆中间载体pMD18-T-RBP1上切下目的基因片段，将目的基因片段与本实验保存的植物过表达载体pCAMBIA3301利用连接酶相连接，构建植物表达载体pCAMBIA3301-ZmRBP1。为进一步研究RBP1的抗冷功能奠定了基础。 关键词：玉米、RBP1基因、植物表达载体、生物信息学分析 PAGE \* MERGEFORMAT PAGE \* MERGEFORMAT I Abstract The reserves of corn are among the forefront of Chinas grain crops, and its production plays a key role in Chinas food security. As the demand for corn continues to increase, conventional breeding methods to cultivate the corn germplasm can not meet the current needs. Therefore, the use of modern biotechnology to improve the traits of maize, to create genes, as well as in the severe conditions of tolerance to become the most urgent needs of the most. As the latitude of the northeast region is relatively high, relatively cold, susceptible to cold air, has also become the corn and other crops the greatest threat, so the development and research of maize cold-related genes become an inevitable trend. The Ran binding protein 1 - Rbp1 protein is a Ran - binding protein and is located in the cytoplasm. The full length of the nucleotide is 1467 bp, which can encode 488 amino acids. In the anti-stress , hormone signal transmission and plant growth and development play an important role . In this study, the cold resistance of maize cold inbred line W9816 was used as the material, and the PCR products were designed by RT-PCR. The obtained RBP1 gene sequence was cloned by extracting RNA and reverse transcription cDNA. Through the bioinformatics analysis of the amino acid sequence of RBP1, the homology wit