摘要
【背景】大肠杆菌中Small RNA EsrE调控琥珀酸脱氢酶的表达并影响细胞生长,对其调控机制的探究有利于加深EsrE对细胞生长影响的认识。【目的】探究大肠杆菌Small RNA EsrE的转录调控机制。【方法】通过双质粒报告系统筛选转录调控因子,并通过凝胶迁移实验(electrophoretic mobilityshiftassay,EMSA)和qRT-PCR研究方法验证转录调控因子。【结果】双质粒报告系统证明RNA聚合酶亚基σ^32(RpoH)上调PesrE,β-羟酰-ACP脱水酶(FabZ)下调PesrE。EMSA结果和体内实验显示RpoH结合PesrE片段,FabZ不结合PesrE片段。【结论】RpoH直接结合启动子序列参与调控,FabZ以其他方式间接参与Small RNA EsrE的转录调控。
[Background]Small RNA EsrE affects cell growth by regulating the expression of succinic acid dehydrogenase in Escherichia coli,the exploration of its regulatory mechanism is conducive to deepening the understanding of EsrE on cell growth.[Objective]To explore the transcriptional regulation mechanism of small RNA EsrE in E.coli.[Methods]Transcriptional regulators were screened by using a dual plasmid reporting system;the interaction between RpoH and PesrE was verified by electrophoretic mobility shift assay(EMSA);qRT-PCR shows effects of the transcriptional regulators on EsrE.[Results]The results of reporting system demonstrated that RpoH up-regulated PesrE,and FabZ down-regulated PesrE.The results of EMSAs showed that RpoH bound the PesrE fragment directly,however FabZ did not.[Conclusion]RpoH is involved in the regulation of esrE transcription by binding the PesrE promoter directly,while FabZ is involved in the transcriptional regulation of EsrE indirectly by other ways.
作者
宋洁
侯兵兵
叶江
吴海珍
张惠展
SONG Jie;HOU Bing-Bing;YE Jiang;WU Hai-Zhen;ZHANG Hui-Zhan(State Key Laboratory of Bioreactor Engineering,School of Biotechnology,East China University of Science and Technology,Shanghai 200237,China)
出处
《微生物学通报》
CAS
CSCD
北大核心
2020年第5期1430-1440,共11页
Microbiology China
基金
国家自然科学基金(31070073,31372550)。