植物转录因子转录活性分析系统开发及验证

Development and Verification of Transcriptional Activity Analysis System for Plant Transcription Factors

转录因子,也称反式作用因子,是调控生物基因转录表达的重要组成部分。其能够与基因启动子区的顺式作用元件(cis-acting element)相互作用,从而激活或抑制基因转录起始,在生物细胞内几乎参与所有生命活动的调控。本文构建了携带转录因子GAL4结合元件的双荧光素酶报告子载体和GAL4-BD融合表达已知转录活性转录因子OsDRAP1和OsMADS57的效应子载体;利用农杆菌介导的烟草瞬时转化体系对该系统进行验证,结果显示,OsDRAP1具有转录激活功能,OsMADS57具有转录抑制功能,表明该系统具有较好的转录活性检测能力,且灵敏性较高,为植物转录因子功能研究和转录相关研究提供了新的技术支撑。

Transcription factor(TF)is also known as trans-acting factor,and it is an important part in the regulation of transcriptional expression of biological genes,which can interact with cis-acting elements in the gene promoter region to activate or inhibit gene transcriptional initiation and participate in the regulation of almost all life activities in biological cells.A dual-luciferase reporter vector with GAL4 cis-element which drives the luciferase reporter gene(LUC)and the effecter vectors with GAL4-BD for fusion expression of OsDRAP1 and OsMADS57 known TF transcriptional activity were constructed,and a transcriptional activity analysis system was verified by using the Agrobacteriummediated transient tobacco transformation system.The results showed that the OsDRAP1 possessed transcriptional activation activity while the OsMADS57 had transcriptional inhibition activity,which indicated that the transcriptional activity analysis system had a strong capability for transcriptional activity detection with high sensitivity and provides new technical support for plant transcription factor function study and transcription related research.