环形电极介导的小麦基因转化  

Wheat Transformation by Electroporation with Ring Electrode

作　　者：梁辉[1,2] 吴方盛 王道文[4] 孙东发[1] 贾旭[1] 

机构地区：[1]华中农业大学植物科学技术学院 [2]中国科学院遗传与发育生物学研究所,北京100101 [3]弗吉尼亚联邦大学生物系 [4]中国科学院遗传与发育生物学研究所

出　　处：《Acta Genetica Sinica》2005年第1期66-71,共6页

基　　金：国家重点实验室开放基金~~

摘　　要：用环形电极电激法有效地将外源DNA导入完整的小麦幼胚组织中。电激的物理参数采用 770V/cm场强、80 0 μF电容、10 0 μg/mL质粒DNA (含有bar和GUS双标记基因 ) ,幼胚被电激 3次。经PCR和Southern杂交分析表明 ,外源基因已稳定整合到小麦基因组中 ,转化频率为 7.5 % ,高于相同处理条件下基因枪法的转化频率(4 2 % )。Electroporation has been used effectively to deliver DNA into the tissue of intact wheat immature embryos.The physical parameters of electroporation were 770 V/cm field strength,800 μF capacitor and 100 μg/mL of plasmid DNA,containing bar and GUS gene.The electroporation was carried out by a portable and permanent ring electrode that can be fitted into the wells of 24-well plates.The samples were pulsed three times.Integration of the introduced genes into the genome of transgenic wheat plants was shown by PCR and Southern blot analysis.Transformation frequency was 7.5%,that is higher than that of the 4.2% frequency by microprojectile bombardment.

关 键 词：电激 环形电极 转化 小麦 幼胚 

分 类 号：S512.1[农业科学—作物学]