芥菜型油菜BjuA03.TTG2基因启动子克隆及表达载体构建  

Clonging of BjuA03.TTG2 Gene Promoter from Brassica juncea and Construction of Its Plant Expression Vectors

作　　者：喻文聪 袁玉辉 李湘 余子怡 伍佳好 向国红 刘显军 YU Wencong;YUAN Yuhui;LI Xiang;YU Ziyi;WU Jiahao;XIANG Guohong;LIU Xianjun(College of Agriculture and Biotechnology,Hunan University of Humanities,Science and Technology,Loudi,Hunan 417000,China)

机构地区：[1]湖南人文科技学院农业与生物技术学院,湖南娄底417000

出　　处：《作物研究》2023年第1期55-61,共7页Crop Research

基　　金：湖南省教育厅科学研究项目(21C0785);湖南人文科技学院研究生科研创新项目(ZSCX2021Y33);湖南省高新技术产业科技创新引领计划项目(2020NK2001)。

摘　　要：为进一步分析芥菜型油菜BjuA03.TTG2启动子在调控该基因表达中的作用,本研究以含有该基因的BAC单克隆质粒DNA为模板,通过普通PCR获得BjuA03.TTG2上游1 839 bp的DNA序列。利用PlantCARE在线分析软件分析BjuA03.TTG2启动子序列,含有许多与应答相关的顺式作用元件,如激素、光、低温等。根据BjuA03.TTG2启动子顺式作用元件的位置设计引物,采用5’端系列缺失分析法,利用PCR方法对BjuA03.TTG2基因上游启动子序列进行特异扩增,分别克隆了1 839、1 497、1 165、863、627、377和297 bp的启动子片段并构建到pCAMBIA1304植物表达载体中。鉴定结果说明7个BjuA03.TTG2启动子不同区段与GUS融合的植物表达载体构建成功。研究结果为进一步分析BjuA03.TTG2启动子的功能作用提供前期基础。In order to further analyze the role of BjuA03.TTG2 promoter in regulating the expression of this gene in Brassica juncea,in this study, we cloned 1 839 bp fragments from the BAC monoclonal plasmid DNA containing the BjuA03.TTG2 by PCR.Using PlantCARE online analysis software to analysis the BjuA03.TTG2 promoter sequence.The result showed that the segment included several cis-acting elements related to response, such as hormone, light, low temperature, etc.Using 5’ end serial deletion analysis method, the specific promoter BjuA03.TTG25’ deletion promoter fragments were amplified by PCR.Promoter with a variation of length 1 839,1 497,1 165,863,627,377 and 297 bp, were ligated with pCAMBIA1304 vector.The identification results showed that plant expression vectors carrying the seven BjuA03.TTG2 promoters with deletion segments driving GUS gene were successfully constructed.These results provided a preliminary basis for further analysis of the function of BjuA03.TTG2 promoter.

关 键 词：芥菜型油菜 BjuA03.TTG2 启动子 植物表达载体构建 

分 类 号：S565.403.2[农业科学—作物学]