RstA/RstB和PhoP/PhoQ系统调控下鼠伤寒沙门氏菌的蛋白质组学研究
发布时间：2021-07-24 09:51
PhoP/PhoQ系统是鼠伤寒沙门氏菌毒力调控中最关键的二元调控系统之一。已知的可以激活PhoP/PhoQ系统的信号包括低浓度的镁离子,酸性pH,以及阳离子抗菌多肽（cAMP） （1,2）。PhoP/PhoQ系统被激活后,可以调节100多个基因的表达（3）,同时PhoP/PhoQ系统对一些其它的二元调控系统,如SsrA/SsrB（4）, PmrA/PmrB（5）和RstA/RstB（6）也有调控作用。有研究表明,沙门氏菌中RstB （RstA/RstB系统中的组氨酸激酶）可以与PhoQ相互作用从而激活PhoP/PhoQ（7）。另外,RstA也可以在低浓度镁离子或酸性pH条件下被PhoP/PhoQ系统激活（8）。这些研究说明在关键调控系统PhoP/PhoQ和了解较少的RstA/RstB系统之间可能存在着关联和互作。通过系统生物学方法研究PhoP/PhoQ系统的工作很多,而对RstA/RstB系统调控网络的系统性研究目前还没有报道。为了更好地解析RstA/RstB系统以及RstA/RstB 与 PhoP/PhoQ之间的关系,我们采用了一个已知的PhoP/PhoQ和RstA/RstB的激活条...
【文章来源】：武汉大学湖北省 211工程院校 985工程院校 教育部直属院校
【文章页数】：153 页
【学位级别】：博士
【文章目录】：
Abstract
中文摘要
Chapter 1:Introduction
1.1 Mass spectrometry-based proteomics
1.1.1 The concept of proteomics
1.1.2 Mass spectrometry basics
1.1.3 Mass spectrometry-based proteomics technique
1.1.4 Quantitative proteomics
1.1.5 Data analysis and bioinformatics tool
1.2 Salmonella pathogenesis
1.2.1 Salmonella overview
1.2.2 Two-component system
1.2.3 The application of proteomics in Salmonella pathogenesis
1.3 Research hypothesis and aims
Chapter 2:Proteomics Analysis of RstA/RstB System in Salmonella Typhimurium
2.1 Introduction
2.2 Materials and methods
2.2.1 Bacterial strains,medium and growth conditions
2.2.2 Protein sample preparation
2.2.3 Two-dimensional electrophoresis methods
2.2.4 Protein digestion
2.2.5 Peptide stable isotope dimethyl labeling
2.2.6 Peptide separation by strong cation exchange(SCX)chromatography
2.2.7 Mass spectrometry analysis and data processing
2.2.8 Bioinformatics analysis
2.2.9 Quantitative real-time PCR(qPCR)analysis
2.2.10 Phenotype analysis
2.3 Results and discussion
2.3.1 Experimental strategies and proteomics workflow
2.3.2 Overview of the proteomics data
2.3.3 Metabolic pathways and function groups regulated by RstA/RstB
2.3.4 RstA/RstB TCS regulation of pyrimidine metabolism
2.3.5 RstA/RstB TCS regulation of iron acquisition
2.3.6 RstA/RstB regulation of surface motility, chemotaxis and invasion
2.4 Conclusion
Chapter 3:Proteomics Analysis of PhoP/PhoQ System in SalmonellaTyphimurium
3.1. Introduction
3.2. Materials and methods
3.2.1. Bacterial strains, medium and growth conditions
3.2.2. Proteins sample preparation
3.2.3 Biofilm phenotype analysis
3.3. Results and discussion
3.3.1 Experimental strategies and proteomics workflow
3.3.2 Overview of the proteomics data
3.3.3 Metabolic pathways and function groups regulated by PhoP/PhoQ system
3.3.4 Network analysis of proteins regulated by PhoP/PhoQ system
3.3.5 PhoP/PhoQ regulates several two-component systems
3.3.6 PhoP/PhoQ regulation of ribosome proteins
3.3.7 PhoP/PhoQ regulates ABC transport proteins
3.3.8 phoPQ mutant effected to biofilm formation
3.4. Conclusion
Chapter 4:Comparative Proteomics Analysis of Salmonella Typhimurium underthe Regulation of RstA/RstB and PhoP/PhoQ Systems
4.1 Introduction
4.2 Materials and Methods
4.3 Results and discussion
4.3.1 Comparative proteomics data overview of the two mutant strains
4.3.2 Comparison of metabolic pathways and functional groups significantly regulated by RstA/RstB and PhoP/PhoQ TCSs
4.3.3 Comparison of common proteins regulated by RstA/RstB and PhoP/PhoQ TCSs
4.4 Conclusion
Chapter 5:Summary and Future Perspective
References
Supplemental S1.80 metabolic pathways of △rstB and △phoPQ strains
Supplemental S2.Changed protein of △rstB strain
Supplemental S3.Changed protein of △phoPQ strain
Research Publications
Acknowledgements
本文编号：3300424
【文章来源】：武汉大学湖北省 211工程院校 985工程院校 教育部直属院校
【文章页数】：153 页
【学位级别】：博士
【文章目录】：
Abstract
中文摘要
Chapter 1:Introduction
1.1 Mass spectrometry-based proteomics
1.1.1 The concept of proteomics
1.1.2 Mass spectrometry basics
1.1.3 Mass spectrometry-based proteomics technique
1.1.4 Quantitative proteomics
1.1.5 Data analysis and bioinformatics tool
1.2 Salmonella pathogenesis
1.2.1 Salmonella overview
1.2.2 Two-component system
1.2.3 The application of proteomics in Salmonella pathogenesis
1.3 Research hypothesis and aims
Chapter 2:Proteomics Analysis of RstA/RstB System in Salmonella Typhimurium
2.1 Introduction
2.2 Materials and methods
2.2.1 Bacterial strains,medium and growth conditions
2.2.2 Protein sample preparation
2.2.3 Two-dimensional electrophoresis methods
2.2.4 Protein digestion
2.2.5 Peptide stable isotope dimethyl labeling
2.2.6 Peptide separation by strong cation exchange(SCX)chromatography
2.2.7 Mass spectrometry analysis and data processing
2.2.8 Bioinformatics analysis
2.2.9 Quantitative real-time PCR(qPCR)analysis
2.2.10 Phenotype analysis
2.3 Results and discussion
2.3.1 Experimental strategies and proteomics workflow
2.3.2 Overview of the proteomics data
2.3.3 Metabolic pathways and function groups regulated by RstA/RstB
2.3.4 RstA/RstB TCS regulation of pyrimidine metabolism
2.3.5 RstA/RstB TCS regulation of iron acquisition
2.3.6 RstA/RstB regulation of surface motility, chemotaxis and invasion
2.4 Conclusion
Chapter 3:Proteomics Analysis of PhoP/PhoQ System in SalmonellaTyphimurium
3.1. Introduction
3.2. Materials and methods
3.2.1. Bacterial strains, medium and growth conditions
3.2.2. Proteins sample preparation
3.2.3 Biofilm phenotype analysis
3.3. Results and discussion
3.3.1 Experimental strategies and proteomics workflow
3.3.2 Overview of the proteomics data
3.3.3 Metabolic pathways and function groups regulated by PhoP/PhoQ system
3.3.4 Network analysis of proteins regulated by PhoP/PhoQ system
3.3.5 PhoP/PhoQ regulates several two-component systems
3.3.6 PhoP/PhoQ regulation of ribosome proteins
3.3.7 PhoP/PhoQ regulates ABC transport proteins
3.3.8 phoPQ mutant effected to biofilm formation
3.4. Conclusion
Chapter 4:Comparative Proteomics Analysis of Salmonella Typhimurium underthe Regulation of RstA/RstB and PhoP/PhoQ Systems
4.1 Introduction
4.2 Materials and Methods
4.3 Results and discussion
4.3.1 Comparative proteomics data overview of the two mutant strains
4.3.2 Comparison of metabolic pathways and functional groups significantly regulated by RstA/RstB and PhoP/PhoQ TCSs
4.3.3 Comparison of common proteins regulated by RstA/RstB and PhoP/PhoQ TCSs
4.4 Conclusion
Chapter 5:Summary and Future Perspective
References
Supplemental S1.80 metabolic pathways of △rstB and △phoPQ strains
Supplemental S2.Changed protein of △rstB strain
Supplemental S3.Changed protein of △phoPQ strain
Research Publications
Acknowledgements
本文编号：3300424
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