利用Ac/Ds转座子系统在水稻中获得无选择标记转基因植株的方法  

Application of Ac/Ds Transposon System to Genetate Marker Gene Free Transgenic Plants in Rice

作　　者：金维正[1] 段瑞君[1] 张帆[1] 陈双燕[1] 吴运荣[1] 吴平[1] 

机构地区：[1]浙江大学生命科学学院

出　　处：《生物工程学报》2003年第6期668-673,共6页Chinese Journal of Biotechnology

基　　金：国家"8 63"专项基金 (No.2 0 0 2AA2Z10 0 3 )~~

摘　　要：获得无选择标记转基因植株是进行重复转基因及消除转基因植株中标记基因潜在危害性的关键。实验采用了Ac Ds转座子系统在水稻 (OryzasativaL .)中进行无hpt选择标记的转基因。将含有目的基因bar的Ds元件和hpt标记基因置于同一个T_DNA中 ,通过农杆菌 (Agrobacteriumtumefaciens)EHA10 5介导将Ac_T_DNA及Ds_T_DNA分别转入到不同的水稻植株 ,再将单拷贝的Ac_T_DNA植株与单拷贝的Ds_T_DNA植株杂交得到同时含有Ac和Ds元件的F1 植株 ,F1 自交产生F2 后代 ,F2 植株中转座后的Ds元件与T_DNA独立分离 ,在总共 10 0株F2 水稻植株中筛选得到 2株只含有Ds元件插入而无hpt标记基因的转基因水稻植株。结果表明 ,利用Ac Ds转座子系统在水稻中获得无选择标记的转基因植株是可行的。It is critical to generate marker gene free transgenic plants for retransformating or eliminating the potential harmfulness of marker gene and its product. In this study, Ac/Ds transposon system was developed for removal of hpt selection marker gene to obtain marker_free transgenic plants in rice (Oryza sativa L.). Ds element containing the interesting gene bar was constructed next to the selection marker gene hpt to get Ds_T_DNA. Rice plants were transformed by Agrobacterium tumefaciens EHA105 containing Ac_T_DNA and Ds_T_DNA respectively. Rice plant containing single copy Ac_T_DNA was crossed with plant containing single copy Ds_T_DNA to obtain the F 1 plant containing both Ac and Ds elements. F 1 plant was self_crossed to produce F 2 progeny in which T_DNA insert and transposed Ds element segregated independently. Two plants contained Ds element but no hpt marker gene in total 100 F 2 plants. The result indicated that Ac/Ds transposon system could be used as a vector system for generating marker gene free transgenic plants in rice.

关 键 词：水稻 无标记基因 转基因技术 Ac/Ds转座子系统 

分 类 号：Q943.2[生物学—植物学]