葡萄Rs基因的克隆序列分析及时空表达特征鉴定
毕业论文关键词：葡萄；Rs基因；克隆；白藜芦醇
Cloning, Sequence Analysis and Characterization of Rs Gene in Vitis vinifera
Abstract：【Objective】To clone the resveratrol synthase gene Rs from Vitis vinifera cv Kyoho, and analyze its sequence and evolution and identify the specificity of the protein in the different developmental stages of fruit 【Method】The total RNA was extracted by CTAB method The Rs gene was cloned by RT-PCR The expression of Rs gene was analyzed by qRT-PCR The sequence of nucleic acid and protein were analyzed by using bioinformatics tools 【Result】The cDNA sequence of the cloned Rs gene was 1247bp in length, and there was an open reading frame (ORF) with a length of 1179bp The gene contained chalcone and stilbene synthase active sites and intact stacclase family Point The results of qRT-PCR showed that the relative expression of Rs gene was the highest in green peel of 25 days after flowering 【Conclusion】The Rs gene was cloned and analyzed, and the specificity of spatiotemporal expression was identified It laid a foundation for further understanding the role of Rs gene in the process of resveratrol metabolism
Key words: Vitis vinifera L , Rs Gene, Clone, Resveratrol
目 录
摘要 1
关键词 1
Abstract 1
Key words 1
1 材料与方法 2
1．1 材料 2
1．1．1 植物材料 2
1．1．2 菌株和载体 2
1．1．3 酶及生化试剂 2
1．1．4 引物与测序 2
1．1．5 培养基 2
1．1．6 主要仪器与设备 2
1．1．7 生物信息学分析 3
1．2 方法 3
1．2．1 葡萄基因组总RNA的提取及cDNA合成 3
1．2．2 目的基因的分子克隆 3
1．2．3 基因的生物信息学分析 3
1．2．4 基因表达特异性分析 3
1．2．5 葡萄皮愈伤组织的诱导与培养 4
1．2．6 细菌转化方法 4
2 结果与分析 4
2．1 葡萄基因组总RNA的提取及cDNA合成 4
2．2 目的基因的克隆 5
2．2．1 基因克隆结果 5
2．2．2 遗传转化结果 5
2．2．3 双酶切结果 6
2．3 基因的生物信息学分析 6
2．3．1 Motif分析 6
2．3．2 基因结构分析 7
2．3．3 蛋白质理化性质及氨基酸组成分析 7
2．3．4 二级结构预测 7