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.
出处
《长江大学学报（自科版）（中旬）》
CAS
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)
关键词
Ran2
超表达载体
RNAI载体
电转化法
Arabidopsis thaliana Ran2 gene
over-expression vector
RNAi vector
electrotransformation
