转录调控研究的方法学进展

Progress in transcriptional studies

基因的表达在多个层次受到严格的控制,其中转录水平的调控是控制基因表达的重要环节之一。近几年,随着分子生物学研究方法的不断发展,转录调控的研究方法学也出现了新的变化。本文综述了基于作者所在课题组对链霉菌次级代谢调控研究过程中优化和改进的转录调控研究方法,如基于SYBR Gold的凝胶阻滞实验,基于荧光标记和毛细管电泳检测的足迹法,基于报告基因的直接调控关系分析等;同时也对研究调控蛋白和靶启动子相互作用的新方法进行了阐述,如表面等离子共振技术和等温滴定量热测定技术等。这些转录调控研究方法的优化和总结,能够帮助研究者开展相关研究。

Gene expression exhibits temporal and spatial patterns to response environmental changes and growth cycle. Gene expression is under strict control at different levels among which control at transcription level is the predominant mode, especially in prokaryotes. In this review, we summarized the new developments of methods used in transcriptional studies, including modifications and improvements of the classic methods, such as gel-shift assay, DNA foot printing, andin vivo reporter system. In addition, we introduced examples to apply new methods, such as surface plasmon resonance （SPR） and isothermal titration calorimetry （ITC） to characterize protein-DNA, ligand-protein, and ligand-protein-DNA interactions. The collection of these methods and their application could guide and accelerate relevant studies.