高粱低磷胁迫响应相关WRKY转录因子克隆和表达分析
毕业论文关键词：高粱；WRKY；磷胁迫；克隆；表达分析
Isolation and Expression Analysis of WRKY transcription factor Respond to low-phosphorous stress in Sorghum
Abstract: Sorghum (Sorghum bicolor L.) is an important food, feed and energy crops. Phosphorous join in most of important physiological process in plants, which is a necessary element for crop growth. It is proteins, nucleic acids, amino acids and chlorophyll composition. However, the content of phosphate in the soil is not enough to satisfy the demand of plant. WRKY transcription factors are a new type transcription regulatory factors found in plants in recent years. In this study, WRKY transcription factors respond to low-phosphorous stress in sorghum was cloned, named SbWRKY2. SbWRKY2 encodes 246 amino acids. Amino acid sequence analysis and homology comparison analysis showed that SbWRKY2 belongs to group Ⅱ of the WRKY transcription factor family. Expression pattern of SbWRKY2 is analyzed under different time in low phosphorus treatment, the results showed that SbWRKY2 were expressed in both roots and shoots. The relative express quantity of SbWRKY2 in the roots is higher than in the shoot’s. Expression of SbWRKY2 induced by low-phosphorus stress and it reached the highest level after treated by low phosphorus for 8 days.
Keywords: sorghum; WRKY; low-phosphate stress; cloning; expression analysis
目 录
摘要：.1
关键词：….1
Abstract1
Keywords.…1
引言… 2
1 材料与方法…..5
1.1 实验材料及处理…..…...5
1.2 主要试剂与仪器…..…...5
1.2.1 主要试剂5
1.2.2 主要仪器...….5
1.3 植物组织总RNA的提取及检测.….….5
1.3.1 植物组织总RNA的提取5
1.3.2 总RNA质量的检测…..6
1.4 cDNA 的合成及检测.….6
1.4.1 cDNA 的合成.6
1.4.2 cDNA 的检测.6
1.5 高粱低磷相关WRKY基因cDNA全长的分离.....7
1.5.1 cDNA全长的分离..7
1.5.2 DNA片段琼脂糖凝胶回收…7
1.6 克隆载体的构建..….8
1.6.1 目的基因与克隆载体的连接…8
1.6.2 大肠杆菌的转化与培养…8
1.6.3 阳性单克隆筛选及测序..8
1.7 生物信息学分析….….….8
1.8 表达特性研究.....9
2 结果与分析...10
2.1 植物组织总RNA与cDNA的检测10
2.2 高粱低磷相关WRKY基因全长的克隆..10
2.3 WRKY基因序列分析及蛋白质特性分析…...11
2.3.1 序列分析及氨基酸预测...11
2.3.2 氨基酸组成及理化性质….…12
2.3.3 氨基酸的同源性分析.…12
2.3.4 蛋白二级结构预测….…13
2.4 SbWRKY2基因的表达分析….14
3 讨论.....15
3.1 SbWRKY2序列特征…...15