利用GFP/RFP双荧光指示载体鉴定特异性启动子功能
Functional Analysis of Specific Promoter Using Vecotors Harboring GFP/RFP Double Fluorescent Marker Genes
投稿时间:2008-05-20  
DOI:  
中文关键词:绿色荧光蛋白, 红色荧光蛋白, 瞬时表达载体, 特异性启动子
英文关键词:green fluorescent proteins, red fluorescent proteins, transient expession vectors, specific promoter
基金项目:国家自然科学基金项目 (No. 30671430) 资助。
作者单位E-mail
尹涛 浙江大学园艺系, 杭州 310029  
秦巧平 浙江林学院, 杭州 311300  
张上隆 浙江大学园艺系, 杭州 310029 shlzhang@zju.edu.cn 
刘敬梅 北京市蔬菜研究中心, 北京 100089  
陈大明 Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, USA  
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中文摘要:
      在基因表达定位或启动子调控模式的研究中, 多以gusA作为报告基因。但由于部分组织中高内源GUS背景活性或转化手段的限制, 使判断基因表达定位或调控时存在很大误差。为了解决上述问题, 本实验将报道基因绿色荧光蛋白(GFP)和红色荧光蛋白(RFP)融合构建双荧光标记瞬时表达载体pBI221-RFP/GFP。该载体以CaMV35S启动子驱动GFP确定转化效率, 通过鉴定阳性个体的红色荧光活性分析目的基因或启动子的表达模式。并通过番茄E8和西瓜AGPL1果实特异启动子验证了该载体在启动子调控模式研究中的应用可行性。结果表明pBI221-RFP/GFP是一个可以在基因和启动子功能验证中应用的高效瞬时表达载体。
英文摘要:
      Most studies related to determining the expression profile of genes and specific promoters used histochemical localization of the reporter gene, gusA. While the histochemical method for visualizing gusA expression suffers from several limitations in the determination of gene expression and location, especially in the tissues with high background acitivty. To solve this problem, a transient expession vector pBI221-RFP/GFP, was constructed using GFP and RFP as double fluorescent marker genes. This vector used CaMV 35S promoter to drive GFP and determine the transforming efficiency. It analyzed expression profile of the target gene and promoter through the RFP activities of the tranformed tissues. Through the specific promoter AGPL1 from watermelon and E8 promoter from tomato, it is resistible to use this vector to study the expression patterns of promoters. Results indicated that the pBI221-RFP/GFP is a very efficient transient expression vector that can be verify the functions of the genes and promoters.
尹涛,秦巧平,张上隆,刘敬梅,陈大明.利用GFP/RFP双荧光指示载体鉴定特异性启动子功能[J].生物工程学报,2008,24(12):2106~2110
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