目录
摘要Ⅱ
关键词Ⅱ
AbstractⅢ
引言
引言
1 材料与方法1
1.1材料 1
1.2方法 1
1.2.1 生物信息学分析1
1.2.2 基因克隆1
1.2.3 酵母双杂交实验2
2 结果与分析3
2.1 GmJAZ17序列分析3
2.2 GmJAZ17蛋白结构分析3
2.3 GmJAZ17基因的启动子元件预测5
2.4 GmJAZ17基因互作蛋白分析6
2.4.1 载体的构建6
2.4.2 自激活与毒性实验7
2.4.3 以pGBKT7GmJAZ17为诱饵筛选酵母文库7
2.4.4 pGBKT7GmJAZ17与pGADT7GmMYC2一对一验证 8
3 讨论 9
4 结论10
致谢10
参考文献12大豆GmJAZ17基因的克隆及互作蛋白鉴定
摘 要
【目的】通过克隆大豆茉莉酸信号通路基因GmJAZ17，并筛选其互作蛋白，从而为深入研究GmJAZ17基因的生物学功能提供参考依据。【方法】以大豆材料Williams 82根部cDNA为模板扩增GmJAZ17基因，应用生物信息学方法分析该基因的理化性质、编码蛋白结构以及启动子区域的顺式作用元件，利用酵母双杂交技术鉴定GmJAZ17的互作蛋白，同时对GmJAZ17和GmMYC2进行一对一验证。【结果】GmJAZ17基因的CDS序列长度为417 bp，编码138个氨基酸，预测其分子量和等电点分别为15.91 kD和9.14。GmJAZ17蛋白具有保守的ZIM和Jas结构域。GmJAZ17的启动子区域包含MYB、MYC转录因子作用元件，此外还包括ABRE、Pbox、ARE等逆境胁迫应答元件。筛选大豆疫霉菌诱导的大豆酵母cDNA文库，结果显示GmJAZ17可以与其他大豆JAZ蛋白形成二聚体，并可与抗真菌的PR5蛋白互作。通过一对一验证证实GmJAZ17与GmMYC2存在互作关系。【结论】GmJAZ17基因与茉莉酸信号紧密相关，可能与其他JAZ蛋白 *51今日免费论文网|www.jxszl.com +Q: @351916072@ 
协同参与茉莉酸信号介导的大豆与大豆疫霉菌的互作。
CLONING AND CHARACTERIZATION OF THE INTERACTING TARGET PROTEIN OF GmJAZ17 IN SOYBEAN
ABSTRACT
【Objectives】In order to provide references for further study on the function of GmJAZ17 gene, we cloned soybean jasmonate signaling pathway gene GmJAZ17 and screened the GmJAZ17interacting proteins. 【Methods】The GmJAZ17 gene was amplified by using the cDNA of Williams 82 root tissue of soybean as template. The physicochemical properties, encoded protein structure and cisacting elements in promoter were analyzed by bioinformatics method. Meanwhile, the GmJAZ17interacting proteins were identified by yeast twohybrid system. Further onetoone verification was analyzed between GmJAZ17 and GmMYC2.【Results】The CDS sequence of GmJAZ17 was 417 bp in length and encoded 138 amino acids. The molecular mass of GmJAZ17 was 15.91 kD, and isoelectric point was 9.14. GmJAZ17 protein contained conserved ZIM and Jas domains. The promoter of GmJAZ17 had MYB and MYC transcription factor elements, and also included stress responsive elements, such as ABRE, Pbox and ARE. Yeast cDNA library was screened which induced by Phytophthora sojae. The results showed that GmJAZ17 could form heterodimerization with other JAZ proteins and interact with PR5 protein. The interaction between GmJAZ17 and GmMYC2 was confirmed by yeast twohybrid. 【Conclusions】GmJAZ17 gene was closely related to jasmonate signal, which may be involved in the interaction between soybean and Phytophthora sojae mediated by JA signal.

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