目录
摘 要 Ⅱ
关键词 Ⅱ
Abstract III
引言
0 引言 1
1 材料与方法 5
1．1 实验材料 5
1．2 技术路线 5
1．3 实验过程 6
1．3．1结构域预测 6
1．3．2表达载体的构建 6
1．3．3烟草瞬时表达实验 7
1．3．4RTPCR检测 8
1．3．5免疫印迹实验 9
2 结果与分析 9
2．1 RGA3的结构域划分 9
2．2 结构域扩增 10
2．3 RGA3全长及各个结构域表达载体的构建 11
2．4 利用烟草瞬时表达系统过表达RGA3全长及各个结构域 12
2．5 RNA水平检测基因表达 13
2．6 RGA3蛋白质免疫印迹 14
3 讨论 14
4 展望 15
致谢 15
参考文献 16
抗病基因类似物RGA3在细胞过敏性反应中的作用
摘 要
小麦的生产对于全球粮食的安全至关重要，但小麦经常遭受白粉病的侵袭，大面积发病时会导致小麦的品质和产量下降。由于白粉菌的无毒基因与小麦抗病基因协同进化，长期使用单一的抗病基因很难对抗新产生的生理小种，因此发掘新的抗白粉病基因是一项长期任务。植物抗病基因编码的功能结构域通常包括：N端结构域、核苷酸结合结构域（NBARC）和Ｃ端富亮氨酸重复序列（LRR）。按照NBSLRR类抗病蛋白N端结构域的差别，将其分为TIRNBSLRR和CCNBSLRR。在禾本科植物中广泛存在着CCNBSLRR类抗病蛋白。本项目对一个来自一粒小麦的抗白粉病基因类似物RGA3的调控机制进行探究。保守结构域预测表明，RGA3编码典型的NBSLRR类抗病蛋白。为明确各个结构域在细胞过敏性反应中的作用，我们将RGA3分为若干长度、编码不同结构域的基因片段，利用农杆菌介导的烟草瞬时表达系统进行分析，发现仅有RGA3全长会引发细胞死亡。本研究为今后发掘更多抗病基因类似物，明确抗病基因的作用机理奠 *51今日免费论文网|www.51jrft.com +Q: @351916072@ 
定基础。
ROLE OF RESISTANCE GENE ANALOG RGA3 IN HYPERSENSITIVE CELL DEATH RESPONSE
ABSTRACT
Wheat production is very important for the global food security, but wheat is often attacked by powdery mildew, which will lead to the decline of wheat quality and yield when a large area of disease occurs. Because the avirulence genes of powdery mildew and the resistance genes of wheat co evolve, it is difficult to use a single resistance gene to resist the new physiological races for a long time, so it is a longterm task to explore new resistance genes of powdery mildew. The functional domains encoded by plant resistance genes usually include:Nterminal domain, nucleotide binding domain (NBARC) and Cterminal leucine rich repeat (LRR). According to the difference of Nterminal domain of NBSLRR, it can be divided into TIRNBSLRR and CCNBSLRR There are CCNBSLRR resistance proteins in Gramineae. This project is to explore the regulatory mechanism of a powdery mildew resistant gene analogue RGA3 from a single wheat. The prediction of conserved domain indicated that RGA3 encoded typical NBSLRR resistance proteins. In order to clarify the role of each domain in cell anaphylaxis, we divided RGA3 into several length, coding different domain gene fragments, and analyzed the Agrobacterium mediated tobacco transient expression system. It was found that only the full length of RGA3 could cause cell death. This study will lay a foundation for exploring more disease resistant gene analogues and clarifying the mechanism of disease resistant genes.

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