可去除选择标记的DREB基因双T-DNA载体共转化玉米

Co-Transformating Maize with Double T-DNA Vector of DREB Gene

通过体外重组构建双T-DNA双元载体pCDMARpWDT-Hyg,选择标记基因hpt(hygromycin phospho-transferase)和抗逆转录因子基因DREB分别位于两个独立的T-DNA。用农杆菌介导法转化玉米胚性愈伤组织,通过在抗性培养基上严格筛选,在获得的再生转化植株中,同时整合hpt基因和DREB基因的共转化率达到26·3%。该转化系统的建立为下一步得到无选择标记的转基因植株奠定基础。此外,本实验还通过gus基因的瞬时表达分析了农杆菌转化玉米的影响因素。

The double T- DNA binary vector pCDMARpWDT- Hyg with two independent T- DNAs, one containing selective marker gene hpt （hygromycin phosphotransferase） and the other containing stress-resistance transcriptional factor gene DREB, was constructed by recombination in vitro and used to transform maize embryonic calli by agrobacterium-mediated method. After strict, selection on resistant medium, the co- transformation ratio of the transformed regenerated plants both by hpt and DREB genes was as high as 26.3 %. The establishment of the double T- DNA vector system provided fundamental data and materials for the development of marker-free transgenic maize plants. Moreover, we discussed the influencing factors to maize transformation by Agrobacterium-mediated method by gus gene transient expression.