目录
摘要Ⅱ
关键词Ⅱ
AbstractⅢ
引言
引言1
1 材料与方法7
1.1 实验材料 7
1.1.1 质粒7
1.1.2 引物7
1.1.3 大肠杆菌7
1.1.4 酵母7
1.2 实验方法 7
1.2.1 构建表达载体7
1.2.2 酵母双杂交10
2 结果与分析12
2.1 酵母筛库结果验证12
2.2 构建Nb40S和NbPRP28基因表达载体12
2.2.1 扩增Nb40S和NbPRP28基因12
2.2.2 筛选阳性克隆13
2.2.3 质粒测序14
2.3 酵母双杂交 17
2.3.1 Nb40S蛋白与Sw5b SD1结构域互作17
2.3.2 NbPRP28蛋白与Sw5b SD1结构域互作18
3 讨论18
致谢20
参考文献20
Sw5b抗病蛋白与Nb40S及NbPRP28蛋白互作研究
摘 要
Sw5b基因从秘鲁番茄（Solanum peruvianum Mill.）中克隆获得，通过识别番茄斑萎病毒（TSWV）移动蛋白（NSm）并介导和激活效应蛋白激发的免疫反应来对抗TSWV的侵染。Sw5b蛋白属于核苷酸结合亮氨酸富集重复受体（Nucleotide binding Leucine rich repeat Receptor, NLR）类免疫受体蛋白，并含有一个额外N端茄科特异结构域（SD），对美洲型正番茄斑萎病毒属（Orthotospoviruses）具有广谱抗性。本研究以模式植物本氏烟（Nicotiana benthamiana）为研究对象，筛选可能参与植物抗病的基因。酵母双杂交（Yeast twohybrid, Y2H）验证表明本氏烟寄主因子Nb40S和NbPRP28与SD互作、同源序列比对发现番茄中存在相应蛋白，为Sw5b免疫受体抗病机制研究提供了线索。作者还对尚待解决的问题作了探讨。
RESEARCH ON T *51今日免费论文网|www.51jrft.com +Q: #351916072# 
HE INTERACTION BETWEEN Sw5b DISEASE RESISTANCE PROTEIN AND Nb40S AND NbPRP28 PROTEIN
ABSTRACT
The Sw5b gene, cloned from Peruvian tomato (Solanum peruvianum Mill.), is a disease resistance gene that confers the resistance to the Tomato spotted wilt virus (TSWV). It mediates recognition of movement protein (NSm) and initiates immune response against TSWV infection. With an additional Nterminal Solanaceaespecific domain (SD), the Sw5b immune receptor is a type of Nucleotide Binding Leucine rich repeat Receptor (NLR) protein and confers a broadspectrum resistance to Tospoviruses. In this study, the plant antivirus background was used to verify the interaction of the host factors Nb40S and NbPRP28 of Nicotiana benthamiana with SD by yeast twohybrid (Y2H). The presence of the corresponding protein in tomato was verified by sequencing to immunize Sw5b. And the author put forward her personal opinions on the issues that need to be resolved in the research.
KEY WORDS: Sw5b; NLR; Orthotospovirus; TSWV; Y2H引言
番茄斑萎病毒（Tomato spotted wilt virus, TSWV）是布尼亚病毒科正番茄斑萎病毒属（Orthotospovirus）典型成员，以西花蓟马为主要传播介体[1]。由于TSWV能感染许多作物，在全世界作物生产中带来重大损失[2, 3]。较早从番茄（Solanum lycopersicum L.）中发现的Sw5是为数不多的已进行商业化探索的抗病资源之一[4]，Steve Tanksley最早将之克隆出来，用于番茄育种。对秘鲁番茄的抗病基因位点测序时，发现了Sw5基因簇[5]。分离其中两个抗性候选基因（RGC）并将其子序列转入烟草植株的实验，最终帮助证明了只有基因拷贝Sw5b能够产生对番茄斑萎病毒的广谱抗性[57]。

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