植物转录因子研究方法及应用

Analytic Methods and Application of Plant Transcription Factors

转录因子功能研究在阐明植物抗逆调控等分子机制中具有重要的作用,依赖于转录因子分析方法的持续创新。本研究从转录因子的发现与鉴定、靶基因与蛋白质互作、蛋白质与蛋白质互作、功能验证4个方面总结了植物转录因子主要分析方法。转录因子的发现与鉴定方法主要包括转录组测序、亚细胞定位、转录因子自激活等;靶基因与蛋白质互作的方法主要有染色质免疫沉淀技术、凝胶电泳迁移、足迹法、酵母单杂交;蛋白质互作的研究方法主要有酵母双杂交、双分子荧光互补、GST-pull down技术;功能验证主要包括突变体表型分析、GUS染色、qRT-PCR等。我们展望了其在热带作物抗逆机制研究中的应用前景,并为植物抗逆的研究提供理论和技术上的指导。

The functional analysis of transcription factors plays an important role in elucidating the regulation mechanism of plant stress resistance, which relay on the continuous innovation of analytic methods of plant transcription factors. We summarize the main analytic methods of plant transcription factors, namely, discovery and identification of transcription factors, the interaction between target genes and proteins, the interaction between proteins and proteins and the functional verification. The methods for the discovery and identification of transcription factors mainly include transcriptome sequencing, subcellular localization, transcription factor autoactivation, etc.. The interaction between target genes and proteins mainly includes chromatin immunoprecipitation, gel electrophoresis migration, footprinting, and yeast one-hybrid. The methods of the interaction between proteins and proteins mainly include yeast two-hybrid, bimolecular fluorescence complementation, GST-pull down technology. Functional verification mainly includes mutant phenotypic analysis, GUS staining, qRT-PCR and so on. Furthermore, the prospects of its application in the study of stress resistance mechanism of tropical crops were reviewed which provide theoretical and technical guidance for the study of plant stress resistance.