拟南芥Ran2基因超表达和RNAi载体的构建  

Construction of Plant Over-expression and RNAi Vector of Ran2 and Its Genetic Transformation of Arabidopsis Thaliana

作　　者：马立安[1] 张忠明[2] 

机构地区：[1]长江大学生命科学学院,湖北荆州434025 [2]华中农业大学农业微生物学国家重点实验室,湖北武汉430070

出　　处：《长江大学学报（自科版）（中旬）》2007年第2期41-44,共4页Journal of Yangtze University(Nature Science Edition)

基　　金：国家自然科学基金项目(30070370)

摘　　要：根据编码拟南芥小GTP结合蛋白的Ran2基因cDNA全序列设计超表达引物和序列内部第397-610bp之间序列设计RNAi引物,以pMD18-T-Ran2为模板,用PCR方法分别扩增出666bp和214bp的片段,分别连接至双元表达载体和RNAi载体上,得到了植物超表达载体pBI-Ran2和RNAi载体Hellsgate2-Ran2,并用电转化法导入农杆菌GV3101菌株中,PCR扩增结果表明所构建的植物超表达载体pBI-Ran2和RNAi载体Hellsgate2-Ran2已导入农杆菌。A 666 bp and a 214 bp fragment were amplified respectively from complete and nucleotides 397 to 610 of the cDNA sequence of Ran2 gene that encodes a Ran2 small GTP-binding protein.The produced fragments were respectively introduced to the binary expressed vector pBI121 to produce pBI-Ran2 and RNAi vector Hellsgate2 to produce Hellsgate2-Ran2.Then pBI-Ran2 and Hellsgate2-Ran2 were respectively mobilized into Agrobacterium tumefaciens strain GV3101 by electrotransformation.The results of PCR amplifications showed that the generated pBI-Ran2 and Hellsgate2-Ran2 were respectively transferred into the Agrobacterium tumefaciens successfully.

关 键 词：Ran2 超表达载体 RNAI载体 电转化法 

分 类 号：Q784[生物学—分子生物学]