籽粒高效特异性表达启动子的克隆与表达分析  

Cloning and Expression Analysis of Grain Specific Expression Promoter

作　　者：王美华[1] 高洁[1] 李玉莲[1] 张淑娟[1] 宋国琦[1] 张荣志[1] 李玮[1] 李吉虎 李根英[1] Wang Meihua;Gao Jie;Li Yulian;Zhang Shujuan;Song Guoqi;Zhang Rongzhi;Li Wei;Li Jihu;Li Genying(Crop Research Institute,Shandong Academy of Agricultural Sciences/Key Laboratory of Wheat Biology and Genetic Improvement in the North Yellow and Huaihe River Valley,Ministry of Agriculture/National Engineering Laboratory for Wheat and Maize,Jinan 250100,China)

机构地区：[1]山东省农业科学院作物研究所/农业部黄淮北部小麦生物学与遗传育种重点实验室/小麦玉米国家工程实验室,山东济南250100

出　　处：《山东农业科学》2021年第5期45-50,共6页Shandong Agricultural Sciences

基　　金：山东省重点研发计划项目(2020CXGC01080513);山东省农业良种工程项目(2019LZGC015);国家重点研发计划项目“主要农作物品质性状形成的分子基础”(2016YFD0100500)。

摘　　要：启动子是调控基因表达的重要元件,本研究从燕麦中克隆了燕麦球蛋白基因的启动子序列,经过比对发现新克隆的启动子序列长960 bp,与参考序列AY795082存在9个SNPs。对启动子组成元件进行分析表明,该启动子含有胚乳特异表达所必须的5个Skn-1(GTCAT)基序;在启动子的正链第622 bp和830 bp处存在胚乳表达相关的顺式调控元件GCN4;在启动子的正链550 bp和负链869 bp处有两个O_(2)位点,曾在玉米中被验证为醇溶蛋白代谢顺式调控元件;在启动子的869 bp处有一个回文结构ACATGTCATCATGT,该序列是胚乳特异表达所必需的。除了上述与胚乳特异性表达有关的元件,该启动子序列中还有许多与逆境响应有关的序列元件。利用上述启动子驱动GUS基因表达试验证明,该启动子启动功能基因表达的强度约为参考序列的5倍,为利用转基因技术改良小麦籽粒性状提供了良好的基因表达驱动元件。Promoter is an important element for gene expression regulation.In this study,the promoter sequence of oat globulin gene was cloned.After alignment,it was found that the length of the new cloned promoter sequence was 960 bp.There were 9 SNPs with reference sequence AY795082.Analysis of the promoter components showed that the promoter contained five Skn-1(GTCAT)motifs necessary for endosperm specific expression.There were cis regulatory elements GCN4 related to endosperm expression at 622 bp and 830 bp of the positive chain of the promoter.There were two O2 sites at 550 bp of positive chain and 869 bp of negative chain of the promoter,which had been verified as the cis regulatory element of gliadin metabolism in maize.There was a palindrome structure ACATGTCATGT at 869 bp,which was necessary for endosperm specific expression.In addition to the above elements related to endosperm specific expression,there were many other elements related to stress response in the promoter sequence.The experiment of GUS gene expression driven by the promoter showed that the intensity of GUS gene expression was about 5 times of the reference sequence,which provided a very good gene expression driving element for improving wheat grain traits by transgenic technology.

关 键 词：启动子 克隆 表达分析 籽粒特异性 

分 类 号：S512.6[农业科学—作物学] Q78[生物学—分子生物学]