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酵母单杂交体系通过筛选cDNA文库可直接得到与靶序列相互作用的蛋白质基因序列,是研究DNA-蛋白质相互作用和转录因子功能的最常用、最有效的方法之一。通过构建与Gal4p的激活结构域融合的表达cDNA文库,利用抗病相关基因启动子区的W-box顺式元件采用酵母单杂交方法对该文库进行了筛选,共得到4个阳性克隆。结果为进一步研究与顺式元件W-box相互作用的WRKY蛋白奠定了基础。
Yeast One-Hybrid System One of the most common and effective methods of studying the function of DNA-protein interactions and transcription factors is to directly obtain the protein gene sequences that interact with target sequences by screening cDNA libraries. By constructing a cDNA library fused with the activation domain of Gal4p, the library was screened by yeast one-hybrid method using the W-box cis-element in the promoter region of the disease-resistant gene. Four positive clones were obtained. The results laid the foundation for the further study of WRKY protein interacting with cis-element W-box.