一种快速检测启动子调控基因表达活性方法的探讨  

Discussion of a Method Which can Speedily Test the Promoter Which Regulates the Gene Expression Activity

作　　者：白斌[1] 张金文[1] 郭志鸿[1] 

机构地区：[1]甘肃省作物遗传改良&种质创新重点实验室

出　　处：《中国农学通报》2005年第7期47-49,共3页Chinese Agricultural Science Bulletin

基　　金：国家自然科学基金(30270843);甘肃省自然科学基金(ZS991-A21-029-N)项目资助。

摘　　要：启动子活性检测是基因工程实验的重要组成部分,利用绿色荧光蛋白(GFP)基因作为报告基因,对rd29A启动子在受体细胞中调控基因表达的活性进行检测,采用基因枪法进行对洋葱表皮细胞的遗传转化,培养24h后在荧光显微镜下观察GFP基因的表达情况,并通过软件测定不同培养条件下GFP基因表达后的荧光强度,能较为准确的鉴定该启动子的活性大小,并达到量化目的。方法简单、省时、可靠,可以作为一种有效的检测启动子活性的方法。Promoter activity detection is the significant component of the genetic engineering. The research utilizes the gene of green fluorescence protein (GFP) as reporting gene, and detects the rd29A promoter which regulates gene expression activities among recipient cell. By particle bombardment to transform the epidermis cells of onion, it observes the gene expression of gfp under the fluorescence microscope after culturing 24h, determining the intensity of fluorescence after gfp is expressed under different culturing conditions so as to diagnose its intensity and achieve the quantization purpose. The method is simple, timesaving and dependable, thus it can be used as a kind of effective measuring in detection of promoter activity.

关 键 词：表达活性 快速检测 绿色荧光蛋白(GFP) RD29A启动子 调控基因表达 GFP基因 启动子活性 组成部分 基因工程 活性检测 报告基因 受体细胞 遗传转化 表皮细胞 基因枪法 镜下观察 荧光显微 荧光强度 培养条件 

分 类 号：Q943.2[生物学—植物学] TQ925[轻工技术与工程—发酵工程]