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  

Strategy for Building Safe Plant Express Vector,an Example of Construct for Expressing Rice Anti-Waxy Gene

作　　者：杨丽君[1,2] 戎益泉[1,2] 张善明[1] 李建粤[1] 

机构地区：[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

出　　处：《分子植物育种》2009年第5期1027-1031,共5页Molecular Plant Breeding

基　　金：上海市农委科技兴农重点攻关项目(农科攻字(2004)第2-17号)资助

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(W axy pro+intron1+anti-W axy)的融合基因单元;而p13AWY-2表达载体含有两个正向排列的W axy pro+intron1+anti-W axy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。In view of the biosafety of genetically modified crops, the idea construct used for plant genetic transformation 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 consumer. 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 developed two express constructs on the basis ofplasmid pCAMBIA1300, named pl3AWY-1 and pI3AWY-2. The expression elements were combined as a fusion unit i.e. Waxy pro＋intronl ＋anti-Waxy and placed in the pl3AWY-1 construct, while the two forward arranged fusion units placed in the pl3AWY-2 construct. In this study, the initial goal we expected would share our thinking and strategies for building safety expression construct and provide case study in the field of rice transgenic research by using rice self promoter and necessary elements without any unsafe DNA element in order to be possible by employing genentic engineering technology for large-scale commercializing use in future.

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 

分 类 号：S511[农业科学—作物学] Q943.2[生物学—植物学]