球孢白僵菌转录因子BbMSN2与其结合基序变体的结合特性分析  

Interactive assays of the Beauveria bassiana transcription factor BbMSN2 and its binding motif variants

作　　者：宋驰[1] 郭凌媛 钱斌 Nemat O.Keyhani 冀宏[1] SONG Chi;GUO Lingyuan;QIAN Bin;Nemat O.Keyhani;JI Hong(College of Biology and Food Engineering,Changshu Institute of Technology,Changshu 215500,Jiangsu,China;Department of Microbiology and Cell Science,Institute of Food and Agricultural Science,University of Florida,Gainesville,32611,Florida,USA)

机构地区：[1]常熟理工学院生物与食品工程学院,江苏常熟215500 [2]Department of Microbiology and Cell Science,Institute of Food and Agricultural Science,University of Florida,Gainesville 32611,Florida,USA

出　　处：《菌物学报》2022年第11期1858-1866,共9页Mycosystema

基　　金：苏州市农业科技创新项目(SNG2021006);常熟市科技发展计划(农业)项目(CN202110)。

摘　　要：球孢白僵菌是一种广谱性杀虫真菌,为了探索其转录因子BbMSN2识别启动子核心序列的能力,本研究外源表达并纯化了BbMSN2蛋白,合成了3个含有不同数量核心序列(AGGGG/CCCCT)的核酸探针和6个核心序列点突变的核酸探针,将BbMSN2蛋白和核酸探针体外结合,通过凝胶迁移实验检测核酸探针及结合蛋白的迁移情况。研究发现,目的蛋白与含有核心序列的核酸探针结合时,核酸探针发生了凝胶迁移现象,其中核心序列数量对凝胶迁移的协同效益不显著。但目的蛋白与核心序列点突变核酸探针结合时,凝胶迁移现象明显减弱。上述结果表明,转录因子BbMSN2可以和含有核心序列核酸探针结合并发生相互作用,且对识别序列具有很强的特异性。本研究为深入探索BbMSN2转录调控机制奠定了试验基础。Beauveria bassiana is a broad spectrum insecticidal fungus.In order to determine the ability of transcription factor BbMSN2 to recognize the promoter binding sites,the BbMSN2 protein was expressed and purified.Three nucleic acid probes with different number of binding site sequences(AGGGG/CCCCT)and six nucleic acid probes with point mutations were synthesized.The protein of BbMSN2 was used to combined with the nucleic acid probes in electrophoretic mobility shift assay.The result showed that BbMSN2 protein could interact with the nucleic acid probes containing the binding sites,but the interaction with point mutation probe was significantly weakened.Moreover,the synergistic effect of the binding sites was not significant.These data indicate that the transcription factor BbMSN2 specifically interact with the nucleic acid probe containing the binding sites.Our study provides foundation for further exploring the transcriptional regulation mechanism of BbMSN2.

关 键 词：转录因子 蛋白表达 凝胶迁移实验 蛋白与核酸相互作用 

分 类 号：S476.12[农业科学—农业昆虫与害虫防治]