酵母双杂交表达载体pGADT7-AtTGA2的构建及酵母菌的转化

[目的] 研究TGA2在植物系统获得抗性中所起的作用,阐明其作用机制.[方法] 通过PCR扩增获得拟南芥中TGA2基因全长CDS序列,克隆至pGADT7酵母表达载体,并转化至AH109酵母菌株中.[结果] 重组质粒通过大肠杆菌转化获得了阳性克隆,经过菌落PCR及单、双酶切验证重组质粒构建成功.将重组质粒又转化到AH109酵母菌中,并在相应筛选培养基上成功获得了转酵母菌成功的阳性克隆.[结论] 为进一步研究TGA2参与水杨酸诱导抗性基因表达的作用机制奠定了良好的基础.

Construction of Yeast Two-hybrid Vectors pGADT7-AtTGA2 and Yeast Transformation

[ Objective ] The aim was to study the role of TGA2 in plant resistance system and clarify its mechanism of action. [ Method] The full length CDS of AtTGA2 was cloned into pGADT7 yeast expression vector and the recombinant plasmid was transformed into AH109 yeast strain. [ Result] Recombinant plasmid obtained positive clones through transformation of escherichia coli, and proven success by colony PCR and single or double enzyme recombinant plasmid. The recombinant plasmid was transformed into AH109 yeast, and the positive clones was obtained success in the corresponding filter medium. [ Conclusion ] The study laid good foundation of further investigation of TGA2 functioning mechanism in the process of resistance gene expression induced by salicylic acid signal.