苦蘵查尔酮合成酶超表达载体的构建及功能研究

摘要类黄酮具有抗氧化、抑制脂质过氧化反应、预防心血管疾病、防癌等的作用。大量研究表明，查尔酮合成酶（CHS）和异构酶（CHI）是类黄酮合成途径中至关重要的两个酶，它们与植物育性、抗逆性密切相关。本试验旨在研究苦蘵查尔酮合成酶（Chalcone synthase，CHS）编码基因的克隆以及在苦蘵的各组织部位中的表达差异。通过TA克隆技术扩增并克隆得到了苦蘵CHS基因的部分序列，采集苦蘵根、茎、叶等，提取各部位总RNA，分析各部位CHS基因的表达差异。结果显示，CHS基因在苦蘵叶部表达量最高。苦蘵的药用部位一般是全株，叶含有大量有益药用成分。本研究克隆的查尔酮合成酶基因，它的表达部位与苦蘵的药用部位是一致的，这说明CHS基因控制的次生代谢途径参与了苦蘵主要药用成分的生物合成。72134

毕业论文关键词：苦蘵；CHS基因；克隆

Flavonoids have functions in antioxidation and the inhibition of lipid peroxidation,prevention of cardiovascular disease and also in cancer prevention。 A large number of studies show that chalcone synthase (CHS) and isomerase (CHI) are two key enzymes in the flavonoid biosynthesis pathway,they are closely related to plant fertility and plant resistance。 The study was designed to detect the cloning and tissue differential expression of chalcone synthetase (CHS) gene in Physalis angulata L。 The sequence of CHS gene was amplified by Original TA Cloning Kit and cloned。 Roots, shoots and leaves of Physalis angulata L。 were collected, and the total RNA of each part was extracted。 The expression level of CHS gene in each part of Physalis angulata L。 was analyzed。 The results showed that the leaf had the highest expression level with a significant difference。 The medicinal parts of Physalis angulata L。 are always the whole plants,the leaves have a lot of useful medicinal ingredients。The cloning of chalcone synthase gene expression location and Physalis angulata L。 medicinal parts consistent,indicating that control CHS secondary metabolic pathways involved in the biosynthesis of Physalis angulata L。 main medicinal ingredients。

Keyword: Physalis angulata L。;CHS gene; Cloning

前言 4

1。研究材料 6

1。1。实验材料 6

1。2。化学试剂 6

1。3。培养基 7

1。4。主要仪器 7

2。研究方法 7

2。1。苦蘵不同部位总RNA提取（Trizol法） 7

2。2。 RNA质量检测 8

2。3。 cDNA模板的合成 8

2。4。 PCR扩增目的基因 9

2。5。目的片段的纯化回收 10

2。6。目的片段与T-载体的连接 11

2。7。转化与蓝白斑筛选 11

2。8。质粒的提取 12

2。9。苦蘵查尔酮合成酶超表达载体的构建及功能研究:http://www.youerw.com/shengwu/lunwen_81995.html