| ABSTRACT | 第7-9页 |
| 中文摘要 | 第10-13页 |
| ABBREVIATION | 第13-15页 |
| Chapter 1: Genome wide analysis of Bacillus thuringiensis reveals partial genescould be potential source of functional toxins | 第15-79页 |
| 1.INTRODUCTION | 第15-45页 |
| 1.1 Bacillus thuringiensis | 第16-29页 |
| 1.1.1 The Bacillus cereus group complex | 第17-19页 |
| 1.1.2 B.thuringiensis virulence factors | 第19-21页 |
| 1.1.3 B.thuringiensis insecticidal crystal protein | 第21-29页 |
| 1.2 Split toxin and partial genes | 第29-31页 |
| 1.3 Structural and functional relationship | 第31-32页 |
| 1.4 Mode of action | 第32-39页 |
| 1.4.1 Solubilisation and proteolytic activati | 第33-34页 |
| 1.4.2 Receptor binding | 第34-35页 |
| 1.4.3 Bravo-Soberon model | 第35-37页 |
| 1.4.4 The "ping-pong" sequential binding model | 第37页 |
| 1.4.5 Zhang-Bulla model | 第37-39页 |
| 1.5 Plant parasite nematodes and B.thuringiensis | 第39-41页 |
| 1.5.1 C.elegans as a model model oranism | 第39-41页 |
| 1.6 Molecular evolution of B.thuringiensis | 第41-44页 |
| 1.7 Aim and objective | 第44-45页 |
| 2.MATERIALS AND METHOD | 第45-55页 |
| 2.1 Bacterial strains | 第45-49页 |
| 2.1.1 Culture medium | 第45页 |
| 2.1.2 Standard polymerase chain reaction (PCR) | 第45-46页 |
| 2.1.3 PCR using thermostable polymerase | 第46页 |
| 2.1.4 Purification of PCR products | 第46页 |
| 2.1.5 Construction of recombinant plasmids | 第46-48页 |
| 2.1.6 Purification of plasmid DNA | 第48页 |
| 2.1.7 Sequencing of plasmid inserts | 第48-49页 |
| 2.2 Culture and maintenance of C.elegans | 第49-53页 |
| 2.2.1 Nematodes bioassays | 第49-50页 |
| 2.2.2 Spore and crystal preparation | 第50-51页 |
| 2.2.3 Protein separation by polyacrylamide gel electrophoresis | 第51页 |
| 2.2.4 Western blot analysis | 第51页 |
| 2.2.5 Dot blot analysis | 第51-52页 |
| 2.2.6 Scanning Electron Microscopy (SEM) | 第52页 |
| 2.2.7 Light microscopy | 第52-53页 |
| 2.3 Bioinformatics methods | 第53-55页 |
| 2.3.1 Mining of Cry protein from B.thuringiensis | 第53页 |
| 2.3.2 Software and databases used | 第53页 |
| 2.3.3 Phylogenetic analysis | 第53-54页 |
| 2.3.4 Statistical analysis | 第54-55页 |
| 3 .RESULTS AND ANALYSIS | 第55-75页 |
| 3.1 Partial genes are widely distributed in the genomes of B.thuringiensis | 第55-60页 |
| 3.2 Bioinformatics analysis of Cry5B-like N-terminal protein | 第60-64页 |
| 3.3 Identification of Cry5B like N-terminal, unable to expression in BMB171 andwild type B.thuringiensis C15 | 第64-66页 |
| 3.4 C-terminal helps in cry stallization of Cry5B-like N-terminal | 第66-69页 |
| 3.5 Hybrid N+C protein is toxic to nematodes | 第69-72页 |
| 3.6 N+C hybrid protein cause pathogenesis through intestine infection | 第72-75页 |
| 4.DISCUSSION AND CONCLUSION | 第75-79页 |
| Chapter 2:Identification of Novel Cry5Fa Crystal protein with Nematicidal Activity | 第79-89页 |
| 1.INTRODUCTION | 第79-82页 |
| 1.1 Plant parasite nematodes a major threat to agriculture | 第79-80页 |
| 1.2 Nematicidal activity of Cry toxins | 第80-81页 |
| 1.2.1 Search for novel cry toxin having wider toxicit | 第80-81页 |
| 1.3 Objective and significance | 第81-82页 |
| 2.MATERIALS AND METHODS | 第82-83页 |
| 3.RESULTS AND ANALYSIS | 第83-89页 |
| 3.1 Genome screening for Novel Cry toxins | 第83-84页 |
| 3.2 Cry5Fa cause the pathogenesis towards C.elegans | 第84-86页 |
| 3.3 Cry5Fa and Cry5Ba showed different activity against mutant C.elegans | 第86-89页 |
| Chapter 3: Hydralysin, Aerolysin like protein as a virulence factor | 第89-98页 |
| 1.INTRODUCTION | 第89-93页 |
| 1.1 Presence of aerolysin like virulance factors in B.thuringiensis | 第90-93页 |
| 2.MATERIALS AND METHODS | 第93页 |
| 3.RESULTS AND ANALYSIS | 第93-97页 |
| 3.1 Bioinformatics analysis of hydralysin | 第93-94页 |
| 3.2 Cloning and Expression of Hydralysin protein | 第94-95页 |
| 3.3 Hydralysin synergistically enhance the activity of Cry5Ba | 第95-97页 |
| 4.CONCLUSION | 第97-98页 |
| REFERENCE | 第98-121页 |
| Published papers and Awards | 第121-123页 |
| APPENDIX | 第123-127页 |
| ACKNOWLEDGEMENT | 第127-129页 |
