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家蚕hsp23.7启动子瞬时表达载体的构建及活性检测

发表时间:2009-06-15  浏览量:1719  下载量:914
全部作者: 张星,胡小龙,王晓娟,谢敏,虞晓华,薛仁宇,曹广力,贡成良
作者单位: 苏州大学医学部基础医学与生物科学学院
摘 要: 为验证家蚕热休克蛋白(heat shock protein,HSP)基因hsp23.7启动子的活性,通过聚合酶链反应(polymerase chain reaction, PCR)扩增得到hsp23.7启动子片段。利用hsp23.7启动子和红色荧光蛋白报告基因(DsRed)构建瞬时表达载体pSK-hsp23.7-DsRed-PolyA,并在家蚕BmN细胞以及家蚕组织中进行瞬时表达研究。结果显示:hsp23.7启动子经热诱导后,能在BmN细胞以及家蚕组织内驱动DsRed基因的表达,表明所克隆的hsp23.7启动子序列具有热休克蛋白基因的启动子活性;序列分析显示,hsp23.7启动子区域二级结构具有复杂的茎环结构,推测其与启动子的功能有关。
关 键 词: 分子生物学;家蚕;热休克蛋白基因启动子;活性检测
Title: Construction of transient expression vector containing hsp23.7 promoter of Bombyx mori and its activity detection
Author: ZHANG Xing, HU Xiaolong, WANG Xiaojuan, XIE Min, YU Xiaohua, XUE Renyu, CAO Guangli, GONG Chengliang
Organization: School of Basic Medicine and Biological Sciences, Medical College of Soochow University
Abstract: In order to verify the activity of the heat shock protein gene promoter (hsp23.7) from silkworm, the hsp23.7 promoter was amplified and cloned into the vector pSK with red fluorescent protein gene (DsRed), then transient expression vector pSK-hsp23.7-DsRed-PolyA was constructed. It was transfected into silkworm BmN cells and silkworm tissues. The result showed that after the hsp23.7 promoter was treated using heat shock, it could induce DsRed gene expression in BmN cells and silkworm tissues. Consequently, the cloned hsp23.7 promoter sequence had the heat shock protein promoter activity. The sequence analysis indicated that secondary structure of hsp23.7 promoter sequence formed a complicated stem-loop structure which could be related with the function of the promoter.
Key words: molecular biology; Bombyx mori; promoter of heat shock protein gene; activity detection
发表期数: 2009年6月第11期
引用格式: 张星,胡小龙,王晓娟,等. 家蚕hsp23.7启动子瞬时表达载体的构建及活性检测[J]. 中国科技论文在线精品论文,2009,2(11):1194-1202.
 
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