甘蓝型油菜BnFAD2-C5基因启动子及内含子在表达水平的功能分析
发布时间：2018-12-30 11:15
【摘要】：富含油酸的菜籽油具有重要的经济价值,使得高油酸育种及油酸形成机制成为热点。油酸脱氢酶基因(FAD2基因)是控制油酸含量的关键酶基因。本文针对Bn FAD2-C5基因展开研究,根据油菜和甘蓝的同源性,克隆了1257 bp启动子序列,利用GUS和GFP作为报告基因分别构建含有不同片段长度的启动子和内含子的缺失载体并转化拟南芥,经GUS染色检测发现 319~ 1 bp为该研究中最小启动子;采用Western blot技术分析启动子和内含子不同区域的功能,发现Bn FAD2-C5启动子区域 1257~ 1020 bp和 319~ 1 bp能够诱导报告基因在转基因拟南芥种子发育中期高效表达,Bn FAD2-C5内含子具有增强启动子转录水平的功能,该功能主要由+631~+1033 bp区域调控。
[Abstract]:Rapeseed oil rich in oleic acid has important economic value, which makes high oleic acid breeding and oleic acid formation mechanism a hot spot. Oleic acid dehydrogenase gene (FAD2 gene) is the key enzyme gene to control oleic acid content. Based on the homology of rape and Brassica napus, the 1257 bp promoter sequence was cloned. GUS and GFP were used as reporter genes to construct promoter and intron deletion vector with different fragment length, and transformed Arabidopsis thaliana. By GUS staining, it was found that 319-1 bp was the smallest promoter in this study. The functions of different regions of promoter and intron were analyzed by Western blot technique. It was found that the promoter region of Bn FAD2-C5 could induce high expression of reporter gene in the middle developmental stage of Arabidopsis thaliana during the development period of transgenic Arabidopsis thaliana. The results showed that the region of Bn FAD2-C5 promoter was 1257 ~ 1020 bp and 319-1 bp. Bn FAD2-C5 intron can enhance the transcriptional level of promoter, which is mainly regulated by 631 ~ 1033 bp region.
【作者单位】： 湖南农业大学农学院/国家油料改良中心湖南分中心;
【基金】：湖南省科技创新项目(CX2013A012) 国家重点基础研究发展规划项目(2015CB150200)资助~~
【分类号】：S565.4
，
本文编号：2395510
[Abstract]:Rapeseed oil rich in oleic acid has important economic value, which makes high oleic acid breeding and oleic acid formation mechanism a hot spot. Oleic acid dehydrogenase gene (FAD2 gene) is the key enzyme gene to control oleic acid content. Based on the homology of rape and Brassica napus, the 1257 bp promoter sequence was cloned. GUS and GFP were used as reporter genes to construct promoter and intron deletion vector with different fragment length, and transformed Arabidopsis thaliana. By GUS staining, it was found that 319-1 bp was the smallest promoter in this study. The functions of different regions of promoter and intron were analyzed by Western blot technique. It was found that the promoter region of Bn FAD2-C5 could induce high expression of reporter gene in the middle developmental stage of Arabidopsis thaliana during the development period of transgenic Arabidopsis thaliana. The results showed that the region of Bn FAD2-C5 promoter was 1257 ~ 1020 bp and 319-1 bp. Bn FAD2-C5 intron can enhance the transcriptional level of promoter, which is mainly regulated by 631 ~ 1033 bp region.
【作者单位】： 湖南农业大学农学院/国家油料改良中心湖南分中心;
【基金】：湖南省科技创新项目(CX2013A012) 国家重点基础研究发展规划项目(2015CB150200)资助~~
【分类号】：S565.4
，
本文编号：2395510
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