根癌农杆菌介导的mMT-IcDNA转化枸杞及其表达的研究  

Study of Expression of mMT I gene by Agrobacterium Mediated Transformation of Lycium barbarum L.

作　　者：赵亚华[1] 何平[1] 高向阳[1] 

机构地区：[1]华南农业大学生物技术学院,广州510642

出　　处：《中国农业科学》2000年第2期92-97,共6页Scientia Agricultura Sinica

摘　　要：报道了小鼠金属硫蛋白 - I c DNA通过根癌农杆菌介导转入枸杞 ,并在枸杞体内表达产生富集锌的转基因枸杞。用带有嵌合 m MT- I c DNA及 ca MV35s启动子的含非致癌性 Ti质粒载体的根癌农杆菌 LBA440 4 ( ca MV35s:m MT- Ic DNA:nos)感染枸杞幼茎外植体 ,在含有Km1 0 0 μg/ml和 Zn2 + 50 μmol/L的选择培养基上筛选转化的愈伤组织 ,并在含 Km50 μg/ml,Zn2 + 1 0 0 μmol/L的分化培养基上得到再生小芽 ,继续转入选择的生根培养基进行生根筛选培养得到完整的再生植株。锌离子富集试验、NPT- 活性检测以及 Southern blot、Western blot的检测结果均表明 ,m MT- Ic DNA已导入枸杞组织并得到表达 ,转 m MT- Ic DNA枸杞对锌离子富集比对照高Young stem segements from Lycium Barbarum L.were infected by A. tumefaciens strain LBA4404 harboring a non oncogenic Ti plasmid derived vector containing a chimaeric gene mMT I cDNA and cauliflower mosaic virus 35s (CaMV35s) promoter as well as nopaline synthase(Non). The calli originated from selective medium (Km50μg/ml+Zn 2+ 100μmol/L) were induced to differentiate into buds and plants regenerated after root induction. The reporter NTP Ⅱ and nopaline activity assay as well as Southern hybridization indicatid that the mMT I cDNA had been integrated into the genome of Lycium Barbarum L. and could be expressed in plants.The Zn 2+ can be bioconcentred by the transgenic Lycium Barbarum L.

关 键 词：金属硫蛋白 枸杞 根癌农杆菌 锌离子富集 转基因 

分 类 号：S188[农业科学—农业基础科学]