摘    要低温作为一种经常发生并且危害严重的环境因素，能够对植物的生长发育产生重大影响。近些年发现拟南芥中的HAP转录因子在抗寒方面起重要作用。低温可诱导HAP转录因子的表达。HAP转录因子能够特异结合启动子中含有CRT/DRE (C-repeat/dehydration re sponsive element )的顺式元件，激活 COR 等基因的表达，从而增强植株抗寒能力，对调控逆境诱导基因的表达具有非常重要的作用。本课题以模式植物拟南芥哥伦比亚生态型植株为实验材料，以含有超表达载体pB7WG2D-HAP的农杆菌进行花序浸染转化，并用除草剂Baste进行阳性植株筛选，得到拟南芥HAP基因超表达株系，利用PCR检测外源基因整合情况。经过筛选，我们最终得到14株突变体，转化率7。78‰。这些转基因株系将用于后续有关HAP基因功能的研究中。89494

毕业论文关键词：拟南芥； HAP基因；突变体；Baste筛选

Low temperature as a frequent occurrence and serious environmental factors, can have a significant impact on plant growth and development。 In recent years, HAP transcription factors in Arabidopsis have been found to play an important role in cold resistance。 Low temperature can induce the expression of HAP transcription factor。 HAP transcription factor can specifically bind to the cis-elements containing the CRT / DRE (C-repeat / dehydration re sponsive element), activate the expression of COR and other genes, thereby enhancing the ability of plant cold resistance, the regulation of stress-induced gene expression Very important role。 In this study, the plants of Arabidopsis thaliana cultivars were used as experimental materials to carry out inflorescence transformation with Agrobacterium tumefaciens containing overexpression vector pB7WG2D-HAP, and positive plants were screened with herbicide Baste to obtain Arabidopsis thaliana HAP gene overexpression The use of PCR to detect foreign gene integration。 After screening, we finally got 14 mutants, the conversion rate of 7。78 ‰。 These transgenic lines will be used in subsequent studies on HAP gene function。

Key word: Arabidopsis thaliana；HAP gene；Mutant；Baste screening

目录

目录 3

1前言 4

1。1 模式植物拟南芥 4

1。2 低温胁迫对植物的影响 4

1。2 植物对低温源Q于W优E尔A论S文R网wwW.yOueRw.com 原文+QQ75201,8766 的响应 4

1。3 本研究的目的意义 5

2 材料与方法 5

2。1 实验材料 5

2。1。1 实验材料 5

2。1。2 材料处理 5

2。1。3 主要试剂 6

2。1。4 主要实验仪器 6

2。1。5 试剂配方 6

2。2 实验方法 7

2。2。1 超表达载体的构建 7

2。2。2 根癌农杆菌LBA4404感受态细胞的转化