您的位置:首页  > 论文页面

DREB基因启动子的克隆及瞬时表达分析

发表时间:2016-08-15  浏览量:1771  下载量:530
全部作者: 陈金焕,张 冲,王厚领,夏新莉,尹伟伦
作者单位: 北京林业大学林木育种国家工程实验室;北京林业大学生物科学与技术学院
摘 要: 以胡杨总DNA为模板克隆,通过PCR技术分离到PeDREB2和PeDREB2L的启动子,并对其核苷酸序列包含的顺式作用元件进行了分析。通过酶切、连接等分析将所克隆的启动子替换pCAMBIA1304中的CaMV35S,成功构建植物表达载体。通过对农杆菌侵染的烟草叶片进行瞬时表达,GUS 组织化学染色和绿色荧光蛋白(green fluorescent protein,GFP)表达检测结果均表明克隆到的序列具有启动活性,能够启动下游标记基因的表达。研究还讨论了启动子序列中的逆境胁迫诱导元件,为植物基因工程提供了一个潜在胁迫诱导型启动子。
关 键 词: 林木育种;胡杨;DREB;启动子活性;瞬时表达
Title: Isolation and transient expression analysis of PeDREB2 promoters from Populus euphratica Oliv.
Author: CHEN Jinhuan, ZHANG Chong, WANG Houling, XIA Xinli, YIN Weilun
Organization: National Engineering Laboratory for Tree Breeding, Beijing Forestry University; College of Biological Sciences and Technology, Beijing Forestry University
Abstract: In this study, the promoters of the PeDREB2 and PeDREB2L were isolated from the total DNA of Populus euphratica Oliv. by PCR and we analyzed the sequence of its nucleotide. The CaMV35S was replaced in pCAMBIA1304 with our cloned promoter by restriction and connection and we successfully constructed a plant expression vector. After infecting the Agrobacterium by transient expression analysis, the result of GUS activity and green fluorescent protein (GFP) imaging in the staining tissue indicated that the cloned sequences have the function of starting the downstream genes’ expression. The stress-induced cis-element component in the promoter region was also discussed and a potential new stress-induced promoter in plant gene engineering will be provided.
Key words: tree breeding; Populus euphratica Oliv.; DREB; promoter activity; transient expression
发表期数: 2016年8月第15期
引用格式: 陈金焕,张 冲,王厚领,等. DREB基因启动子的克隆及瞬时表达分析[J]. 中国科技论文在线精品论文,2016,9(15):1507-1513.
 
0 评论数 0
暂无评论
友情链接