【摘要】：富含油酸的菜籽油具有重要的經(jīng)濟(jì)價(jià)值,使得高油酸育種及油酸形成機(jī)制成為熱點(diǎn)。油酸脫氫酶基因(FAD2基因)是控制油酸含量的關(guān)鍵酶基因。本文針對(duì)Bn FAD2-C5基因展開(kāi)研究,根據(jù)油菜和甘藍(lán)的同源性,克隆了1257 bp啟動(dòng)子序列,利用GUS和GFP作為報(bào)告基因分別構(gòu)建含有不同片段長(zhǎng)度的啟動(dòng)子和內(nèi)含子的缺失載體并轉(zhuǎn)化擬南芥,經(jīng)GUS染色檢測(cè)發(fā)現(xiàn) 319~ 1 bp為該研究中最小啟動(dòng)子;采用Western blot技術(shù)分析啟動(dòng)子和內(nèi)含子不同區(qū)域的功能,發(fā)現(xiàn)Bn FAD2-C5啟動(dòng)子區(qū)域 1257~ 1020 bp和 319~ 1 bp能夠誘導(dǎo)報(bào)告基因在轉(zhuǎn)基因擬南芥種子發(fā)育中期高效表達(dá),Bn FAD2-C5內(nèi)含子具有增強(qiáng)啟動(dòng)子轉(zhuǎn)錄水平的功能,該功能主要由+631~+1033 bp區(qū)域調(diào)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(nóng)業(yè)大學(xué)農(nóng)學(xué)院/國(guó)家油料改良中心湖南分中心;
【基金】：湖南省科技創(chuàng)新項(xiàng)目(CX2013A012) 國(guó)家重點(diǎn)基礎(chǔ)研究發(fā)展規(guī)劃項(xiàng)目(2015CB150200)資助~~
【分類號(hào)】：S565.4
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本文編號(hào)：2395510