声明
TABLE OF CONTENTS
LIST OF TABLES
LIST OF FIGURES
LIST OF ABBREVIATIONS
摘要
ABSTRACT
CHAPTER 1 INTRODUCTION AND LITERATURE REVIEW
1 Epidemiology and pathogenesis of Streptococcus suis
1．1Introduction of S． suis(SS)
1．2Diseases caused by S． suis(SS)
1．3Epidemiology of Streptococcus suis infection
1．4Virulence factors of S． suis(SS)
2 Introduction to ATP-dependent proteases
2．1The AAA+ Superfamily of Proteins
2．2Stress tolerance of ATP-dependent proteases
2．3Control of clp expression
2．4Clp proteins affecting virulence of pathogens
Reference
CHAPTER 2 ClpP IS REQUIRED FOR THE STRESS TOLERANCES AND VIRULENCE IN STREPTOCOCCUS SUIS SEROTYPE 2
1 Material and methods
1．1Bacterial strains，culture conditions，plasmids，and host cell lines
1．2DNA manipulations
1．3Recombinent DNA technique
1．4Growth characteristics and genetic stability of mutant strains
1．5Grams stain assay
1．6Biofilm plate assay
1．7Survival assays of oxidative(H2O2)，acidic pH and osmotic(NaCl)condition in SS2
1．8Adhesion and invasion assays
1．10Animal infection model
1．11RNA extraction and qRT-PCR analysis
1．12Bioinformatics and statistical data analysis
2 Results
2．2ClpP is required for the optimal growth of SS2 during heat stress
2．3Deletion of clpP caused a significant deficiency in bacterial chain and biofilm formation
2．4Deletion of clpP attenuated oxidative and acidic tolerance，but not sensitivity to osmotic stress
2．5ClpP in SS2 contribute to antiphagocytosis against RAW264．7 cells and adhesion to HEp-2 cells
2．6Effect of clpP deletion during systemic infection in vivo
2．7Expression profiling of virulence gene expression via qRT-PCR
3 Discussion
4 Conclusion
References
CHAPTER 3 ROLE OF ClpX IN SS2 STRESS TOLERANCE AND VIRULENCE
1 Materials and methods
1．1Bacterial strains，culture conditions，plasmids and host cell lines
1．2DNA manipulations
1．3Recombinant DNA techniques
1．4Growth characteristics and genetic stability of mutant strains
1．5Grams stain assay
1．6Survival assays of oxidative(H2O2)，acidic pH and osmotic(NaCl)condition in SS2
1．7Cell experiments
1．8Animal infection model
1．9RNA extraction and qRT-PCR analysis
1．10 Bioinformatics and statistical data analysis
2 Results
2．2ClpX is required for the optimal growth in cold stress of SS2
2．3 clpX deletion caused a significance deficiency in bacterial chain’s truncation
2．4Inactivation of ClpX attenuated oxidative and acidic pH tolerance，but not sensitivity to osmotic stress
2．5ClpX in SS2 contribute to antiphagocytosis against RAW264．7 cells and adhesion to HEp-2 cells
2．6ClpX plays key roles in the pathogenic processes of SS2
2．7Expression profiling of virulence gene expression via qRT-PCR
3 Discussion
4 Conclusion
References
CHAPTER 4 THE ROLES OF FILAMENTATION TEMPERATURE SENSITIVE H(FtsH)IN THE SURVIVAL OF SS2 UNDER STRESSES AND THE VIRULENCE IN BALB／C MICE
1 Materials and methods
1．1Bacterial strains，culture conditions，plasmids，and host cell lines
1．2DNA manipulations
1．3Recombinant DNA techniques
1．4Growth characteristics and genetic stability of mutant strains
1．5Biofilm plate assay
1．6Survival assays under oxidative(H2O2)，acidic pH and osmotic(NaCl)conditions in SS2
1．7Adhesion and invasion assays
1．8Phagocytosis assay
1．10Animal infection model
1．11Electrophoretic Mobility Shift Assay(EMSA)
1．12RNA extraction and qRT-PCR analysis
1．13Bioinformatics and statistical data analysis
2 Results
2．1Identification of FtsH homologs in SS2
2．2ftsH deletion affects growth at a high temperature
2．3Deletion of ftsH reduced biofilm formation
2．4Deletion of ftsH attenuated oxidative，acidic tolerance and osmotic stress
2．5FtsH in SS2 contribute to antiphagocytosis against RAW264．7 cells and adhesion to HEp-2 cells
2．6Effect of ftsH mutations during systemic infection in vivo
2．7Purified CtsR repressor protein can not bind to the promoter regions of ftsH
2．8Heat shock induces ftsH expression in S．suis
3 Discussion
4 Conclusion
References
CHAPTER 5 THE CtsR REGULATED THE EXPRESSION OF clpP IN SS2
1Materials and methods
1．1Bacterial strains，culture conditions，and plasmids
1．2DNA manipulations
1．3Recombinant DNA techniques
1．4Survival assays of oxidative and temperature condition in SS2
1．5Electrophoretic mobility shift assay (EMSA)
1．7Animal infection model
1．8RNA extraction and qRT-PCR analysis
1．9Bioinformatics and statistical data analysis
2 Results
2．1Identification of the CtsR(class three stress gene repressor)in SS2
2．2Role of CtsR during indunction of stress
2．3 Prediction and transcriptional analysis of putative CtsR regulon members
2．4Purified CtsR binds specifically to the promoter regions of clpP
2．5The role of CtsR in the pathogenicity of S．suis
3 Discussion
4 Conclusion
References
GENERAL CONCLUSIONS
PUBLICATIONS
INNOVATIONS
ACKNOWLEDGEMENTS
南京农业大学;
