【摘要】：富含油酸的菜籽油具有重要的經濟價值,使得高油酸育種及油酸形成機制成為熱點。油酸脫氫酶基因(FAD2基因)是控制油酸含量的關鍵酶基因。本文針對Bn FAD2-C5基因展開研究,根據油菜和甘藍的同源性,克隆了1257 bp啟動子序列,利用GUS和GFP作為報告基因分別構建含有不同片段長度的啟動子和內含子的缺失載體并轉化擬南芥,經GUS染色檢測發(fā)現 319~ 1 bp為該研究中最小啟動子;采用Western blot技術分析啟動子和內含子不同區(qū)域的功能,發(fā)現Bn FAD2-C5啟動子區(qū)域 1257~ 1020 bp和 319~ 1 bp能夠誘導報告基因在轉基因擬南芥種子發(fā)育中期高效表達,Bn FAD2-C5內含子具有增強啟動子轉錄水平的功能,該功能主要由+631~+1033 bp區(qū)域調控。
[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.
【作者單位】： 湖南農業(yè)大學農學院/國家油料改良中心湖南分中心;
【基金】：湖南省科技創(chuàng)新項目(CX2013A012) 國家重點基礎研究發(fā)展規(guī)劃項目(2015CB150200)資助~~
【分類號】：S565.4
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本文編號：2395510