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草莓中农杆菌tdna插入位点特征研究【字数:8905】

2024-11-03 13:05编辑: www.jxszl.com景先生毕设

目录
摘 要Ⅱ
关键词Ⅱ
AbstractⅢ
引言
引言1
1 材料与方法3
1.1实验植株材料和培养条件 3
1.2 CTAB法提取植物基因组 3
1.3 用染色体步移技术获得TDNA插入片段 3
1.4 确定TDNA插入位点 4
1.4.1 获取森林草莓的DNA序列 4
1.4.2 侧翼序列的预处理和注释选择4
1.4.3 进行多重序列比对确定插入位点 4
1.5 GO和KEGG Pathway显著富集分析 4
1.6 基因组DNA制备和BSseq检测DNA甲基化情况 4
1.6.1 测定基因组DNA的降解、污染和浓度 4
1.6.2 文库准备和定量 4
1.6.3 数据整合分析DNA甲基化情况 5
1.7 插入位点区域GC含量和甲基化频率与TDNA插入位点的相关性 5
1.8 使用Monte Carlo method模拟TDNA的随机插入 5
2 结果与分析6
2.1 TDNA在染色体和基因组水平的插入分布 6
2.2 TDNA插入位点与碱基对GC含量关系 7
2.3 TDNA插入位点数量与基因表达水平的关系 8
2.4 TDNA插入位点与基因甲基化的关系 9
3 讨论10
参考文献11
致谢13
草莓中农杆菌TDNA插入位点特征研究
摘 要
农杆菌介导的转基因方法是目前植物遗传转化的重要方法之一。该方法通过农杆菌将外源TDNA转移到植物细胞中,然后整合到植物基因组实现转基因,所以TDNA整合到基因组的效率直接影响植物的转化效率。而有关农杆菌介导转化时插入位置的研究表明,TDNA的整合位点可能不是随机的,有其特定规律和一定偏好性。但目前对草莓中农杆菌介导的TDNA插入位点特征的研究还较少,因此本实验以“草莓中农杆菌介导转化的TDNA插入位点特征研究”为主题来开展研究。首先准备好合适的实验材料进行农杆菌介导,接着用 *51今日免费论文网|www.51jrft.com +Q: &351916072
染色体步移技术获取TDNA插入片段,之后从网上数据库完成的森林草莓(Fragaria vesca)测序序列用于比对,最后通过多种测试方法和数据统计和制图分析软件进行数据分析,进而得出TDNA在基因组和染色体水平的插入分布,并探究了TDNA插入位点与基因表达水平、GC含量及基因甲基化之间的相关性,以揭示草莓中TDNA插入的偏好性及可能的机制。本研究目的为对草莓中农杆菌介导的TDNA插入位点的特征进行分析,为提高草莓遗传效率提供理论基础。
IDENTIFICATION AND CHARACTERIZATION OF TDNA INSERTION SITES CREATED BY AGROBACTERIUMMEDIATED TRANSFORMATION IN STRAWBERRY
ABSTRACT
Agrobacteriummediated transformation is one of the most important methods of plant genetic transformation. In this method, exogenous TDNA was transferred into plant cells by Agrobacteriummediated transformation, and then integrated into the plant genome,so the efficiency of TDNA integration into the genome directly affects the efficiency of plant transformation. Studies of TDNA insertion sites by Agrobacteriummediated transformation suggest that the integration sites of TDNA may not be random, have their specific regularity and preferences.At present, there are few studies on the characteristics of TDNA insertion site by Agrobacteriummediated transformation in strawberries. Therefore, this study carried out the experiment with "identification and characterization of TDNA insertion sites created by Agrobacteriummediated transformation in strawberry" as the research topic.First, appropriate experimental materials were prepared for Agrobacteriummediated transformation, and then the experiment of strawberry plants with chromosome walking technologies for their insert TDNA fragment.the database of the wood land strawberry(Fragaria vesca) from the Internet is used to compare the sequence , finally through a variety of testing methods and statistics and graphics analysis software for final data analysis ,we can get T DNA insert distribution of chromosomes, and the correlation between the TDNA insertion site and gene expression level, GC content or gene methylation in the genome,to reveal the preference of TDNA insertion in strawberry and its possible mechanism. This study provides a theoretical basis for improving the genetic efficiency of strawberries by analyzing the characteristics of TDNA insertion sites by Agrobacteriummediated transformation in strawberries.

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