植物安全性表达载体的构建策略以省略达水稻反义蜡质基因的载体构建为例杨丽君.pdfVIP

分子植物育种，2009 年，第7 卷，第5 期，第1027-1031 页 Molecular Plant Breeding, 2009, Vol.7, No.5, 1027-1031 新思路、新技术、新方法 Novel Thinking Technology 植物安全性表达载体的构建策略：以表达水稻反义蜡质基因的载体构建 为例 1,2 1,2 1 1* 杨丽君 戎益泉 张善明 李建粤 1 上海师范大学生命与环境科学学院, 上海, 200234; 2 上海医药高等专科学校, 上海, 201318 * 通讯作者, lijianyue01@ 摘 要 基于转基因作物的安全性考虑，在用于转化的表达载体上除了含有目的基因以及控制该目的基因表 达的启动子和终止子外，最好不存在其它有可能存在安全性争议的DNA 序列，由此培育的转基因植物可能将 更易于被消费者所接受。本研究以pCAMBIA1300 载体为基础，基因操作去除pCAMBIA1300 质粒上的潮霉 素抗性基因和花椰菜花叶病毒的35S 启动子序列，构建了两种只含有水稻蜡质基因启动子引导蜡质基因反义 片段的表达载体，p13AWY-1 和p13AWY-2。其中p13AWY-1 表达载体含有一个由水稻蜡质基因启动子、第 一内含子、反义蜡质基因(Waxy pro+intron1+anti-Waxy) 的融合基因单元；而p13AWY-2 表达载体含有两个正 向排列的Waxy pro+intron1+anti-Waxy 融合基因单元。我们通过构建水稻反义蜡质基因安全性表达载体，试图 为植物安全性表达载体的构建提供一种思路，为今后大规模商业化采用转基因技术改良农作物遗传特性提供 安全的转基因方法。 关键词 转基因植物, 植物表达载体, 生物安全性, 35S 启动子, 抗生素标记, 水稻 Strategy for Building Safe Plant Express Vector, an Example of Construct for Expressing Rice Anti-Waxy Gene Yang Lijun 1,2 Rong Yiquan 1,2 Zhang Shanming 1 Li Jianyue 1* 1 College of Life and Environment Science, Shanghai Normal University, Shanghai, 200234; 2 Shanghai Institute of Health Sciences, Shanghai, 201318 * Corresponding author, lijianyue01@ DOI: 10.3969/mpb.007.001027 Abstract In view of the biosafety of genetically modified crops, the idea construct used for plant genetic trans- formation should only contain expression elements such as target gene, promoter and terminator, which would be necessary for expressing the target gene. And also it is strong recommended that the biosafe construct should have no any uncertain element and exogenous DNA sequence such as antibiotics gene, 35S promoter etc. It is no doubt that the transgenic products developed by using safety expression construct might be increasing accepted by con- sumer. In this study, we put forward a strategy for building a kind of safety plant expression construct by removing both of hygromycin resistant gene and the cauliflower mosaic virus 35S promoter, and then we d