生物工程专业毕业论文--红豆杉MYB-1基因的克隆及其表达产物的亚细胞定位分析.docVIP

红豆杉MYB-1基因的克隆及其表达产物的亚细胞定位分析 摘 要 克隆红豆杉叶片细胞中的MYB-1转录因子基因，对基因序列进行初步的生物信息学分析，再通过构建亚细胞定位载体，对基因的表达产物进行定位分析。 本研究首先提取红豆杉细胞的总RNA后，RT-PCR得到cDNA第一链，经检验，可以用作模板扩增基因。根据MYB-1基因的ORF设计引物，扩增MYB-1基因,得到1668bp的完整开放阅读框，编码555个氨基酸。然后对测序结果进行结构域分析，表明其存在DNA结合结构域，并对其二级结构进行了预测，将它与多种的MYB家族基因进行多序列比对，分析其保守结构域，进一步构建相应的系统进化树。由测序结果设计特异性引物，并成功构建了亚细胞定位载体 ，最后，基因枪转化洋葱表皮进行定位分析，证实MYB-1蛋白在核内定位。为进一步分析研究其详细功能奠定了基础。 关键词： 红豆杉 MYB-1转录因子 进化树分析 亚细胞定位 Cloning Of MYB-1 Gene From Taxus And The Subcellular Localization Of Its Expression Product Abstract Cloning MYB-1 transcription factor genes from Taxus leaf cells, gene sequences preliminary bioinformatics analysis, and then by constructing the subcellular localization vector for analysis the localization of gene expression product.In this study, total RNA extracted from Taxus cells, first strand cDNA obtained through RT-PCR. By inspection, it can be used as a template to amplified genes. According the ORF of MYB-1 gene, primers designed to PCR amplification MYB-1 gene. We achieved a complete open reading frame of 1668bp of objectve fragment, encoding 555 amino acids. Then analysis structural domain of sequencing results, indicating that the existence of DNA binding domain, and its secondary structure was predicted, through alignment with a variety of multiple sequence, and analysis the conserved domain, then build corresponding phylogenetic tree further. By the sequence specific primers were designed and successfully constructed subcellular localization vector, and finally, insert into the onion epidermal by gene gun, positioning analysis confirmed MYB-1 protein in nuclear localization. This result laid a sound foundation for the next step that the analysis of its function.Key words: Taxus MYB-1 transcription factor phylogenetic trees subcellular localization目 录 HYPERLINK \l _Toc199479692 摘要﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍Ⅰ HYPERLINK \l _Toc199479692 Abstract ﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍﹍Ⅱ