小麦TaMyb2基因转化拟南芥表达载体的构建与遗传转化  

Construction and Genetic Transformation of Arabidopsis thaliana Expression Vector of Wheat TaMyb2

作　　者：王爱萍[1,2] 景蕊莲[2] 杨武德[1] 董琦[1] 

机构地区：[1]山西农业大学农学院,太谷030801 [2]中国农业科学院作物科学研究所/农作物基因资源与基因改良国家重大科学工程/农业部作物种质资源利用重点开放实验室,北京100081

出　　处：《应用与环境生物学报》2008年第6期750-752,共3页Chinese Journal of Applied and Environmental Biology

基　　金：山西省青年科技研究基金项目(No.2008021044);国家“973”重点基础研究发展计划项目(No.2004CB117202)资助~~

摘　　要：采用双酶切方法构建了小麦TaMyb2基因转化拟南芥表达载体pCHF3-35Sp-TaMyb2-Ⅰ-NOSter和pCHF3-35Sp-TaMyb2-Ⅱ-NOSter,并通过农杆菌GV3101介导,用渗透法将TaMyb2-Ⅰ和TaMyb2-Ⅱ基因转入拟南芥中.结果表明,TaMyb2-Ⅰ、TaMyb2-Ⅱ和表达载体pCHF3分别被KpnⅠ和PstⅠ成功酶切为837bp、840bp和10.4kb的目标条带;分别将pCHF3-35Sp-TaMyb2-Ⅰ-NOSter和pCHF3-35Sp-TaMyb2-Ⅱ-NOSter转入农杆菌GV310l中,在提取的农杆菌质粒DNA中分别扩增出了837bp和840bp目标条带,说明可用于拟南芥的转化;在含Kan(50mg/L)的1/2MS培养基上筛选转基因当代拟南芥植株的种子(T0),获得了T1代抗性植株,转化率约为0.1%.Arabidopsis thaliana expression vectors pCHF3-35Sp-TaMyb2-Ⅰ -NOSter and pCHF3-35Sp-TaMyb2-Ⅱ-NOSter constructed by double restriction endonucleases and genes TaMyb2-Ⅰ, TaMyb2-Ⅱ were transformed into A. thaliana mediated by Agrobacterium GV3101 by infiltration. The results showed that TaMyb2-Ⅰ, TaMyb2-Ⅱ and expression vector pCHF3 were successfully digested by Kpn Ⅰ and PstⅠ, and the lengths of target segments were 837 bp, 840 bp and 10.4 kb, respectively; pCHF3-35Sp-TaMyb2-Ⅰ-NOSter, pCHF3-35Sp-TaMyb2-Ⅱ-NOSter and the empty vector pCHF3 were transformed into Agrobacterium GV3101, respectively. The target segments 837 bp and 840 bp were amplified from Agrobacterium plasmid DNA, respectively. It showed that Agrobacterium was suitable for the next transformation experiment. T0 resistant plant was obtained from the 1/2 MS medium comprising Kan （50 mg/L） by screening the seeds （T0） of the transformed Arabidopsis plants. The transformation rate was about 0.1%. Fig 3, Ref 14

关 键 词：TaMyb2-Ⅰ TaMyb2-Ⅱ 拟南芥 表达载休 遗传转化 

分 类 号：S512.1[农业科学—作物学]