| 中文摘要 | 第5-7页 |
| Abstract | 第7-8页 |
| Abbreviations | 第12-13页 |
| List of Figures | 第13-15页 |
| List of Tables | 第15-16页 |
| CHAPTER Ⅰ Literature review | 第16-38页 |
| 1.1 Discovery and impact of Southern rice black-streaked dwarf disease | 第16-18页 |
| 1.2 Genome and structure of SRBSDV | 第18-20页 |
| 1.3 The vector of SRBSDV-Sogatella furcifera | 第20-23页 |
| 1.3.1 Vector taxonomy and characteristics | 第20-22页 |
| 1.3.2 Vector's life cycle | 第22-23页 |
| 1.4 Characteristics of SRBSDV-WBPH-plant host interaction | 第23-24页 |
| 1.5 Plant virus transmission by insect vector | 第24-31页 |
| 1.5.1 Mode of virus transmission by insect vector | 第25-27页 |
| 1.5.2 Virus determinants and vector's components involved in virus transmission | 第27-31页 |
| 1.6 Detection of Protein-protein interactions | 第31-37页 |
| 1.6.1 The Split-ubiquitin yeast two hybrid system | 第32-34页 |
| 1.6.2 Chemiluminescent Coimmunoprecipition(Co-Ip) | 第34-37页 |
| 1.7 Objectives | 第37-38页 |
| CHAPTER Ⅱ Investigation of protein interactors by yeast two hybrid system | 第38-62页 |
| 2.1 Introduction | 第38-39页 |
| 2.2 Material and Methods | 第39-53页 |
| 2.2.1 Virus and insect vector sources | 第39页 |
| 2.2.2 Construction of Sogatella furcifera cDNA library fusion vector | 第39-41页 |
| 2.2.3 Construction of SRBSDVP7-1 bait fusion vector | 第41-47页 |
| 2.2.4 Transformation of P7-1 bait fusion vector into yeast | 第47-48页 |
| 2.2.5 Preparation of yeast cultures and protein extraction | 第48页 |
| 2.2.6 SDS/PAGE and Western blotting | 第48-49页 |
| 2.2.7 Verification of pDHB1-P7-1 expression | 第49-50页 |
| 2.2.8 Optimization of basic screening conditions | 第50-51页 |
| 2.2.9 Library screen and selection of interactor | 第51-52页 |
| 2.2.10 Annotation and gene ontology(GO)classification | 第52页 |
| 2.2.11 Confirmation of positive interactors | 第52-53页 |
| 2.3. Results | 第53-61页 |
| 2.3.1 Construction of S.furcifera cDNA library | 第53-54页 |
| 2.3.2 Construction of P7-1 bait fusion vector | 第54-55页 |
| 2.3.3 Verification of pDHB1-P7-1 expression by Western blot | 第55-56页 |
| 2.3.4 Veilfication of bait fusion protein correct expression | 第56-57页 |
| 2.3.5 Non-self activation test and optimization of library screen | 第57页 |
| 2.3.6 Investigation of interactors by library screen | 第57-58页 |
| 2.3.7 Annotation of bioinformatics and gene ontology(GO)classification | 第58-60页 |
| 2.3.8 Confirmation of positive interactors in yeast | 第60-61页 |
| 2.4. Discussion | 第61-62页 |
| CHAPTER Ⅲ Identification of protein interactors by Chemiluminescent Co-IP system | 第62-72页 |
| 3.1 Introduction | 第62页 |
| 3.2 Materials and Methods | 第62-67页 |
| 3.2.1 Cloning of AcGFP1-bait and pProLabel-prey | 第62-64页 |
| 3.2.2 Cotransfection of AcGFPl-Bait and ProLabel-Prey into Mammalian Cells | 第64页 |
| 3.2.3 Verification of AcGFP1-Bait expression and preparation of cotransfected cells | 第64页 |
| 3.2.4 Verification of ProLabel-prey expression | 第64-65页 |
| 3.2.5 Chemiluminescent CO-IP assay | 第65-67页 |
| 3.3 Results | 第67-71页 |
| 3.3.1 Construction of bait and prey fusion vectors | 第67-68页 |
| 3.3.2 Verification of AcGFP1-bait and ProLabelC-Prey expression | 第68-70页 |
| 3.3.3 Chemiluminescent coimmunoprecepitation(CO-IP)assay | 第70-71页 |
| 3.4 Discussion | 第71-72页 |
| CHAPTER Ⅳ Identification of mRNA expression level by qRT-PCR | 第72-78页 |
| 4.1 Introduction | 第72页 |
| 4.2 Materials and Methods | 第72-74页 |
| 4.2.1. Gene expression analysis by qRT-PCR | 第72-74页 |
| 4.3. Results | 第74-77页 |
| 4.3.1 Gene expression analysis in different growth stages of S.furcifera | 第74页 |
| 4.3.2 Gene expression analysis in different organs of S.furcifera | 第74-77页 |
| 4.4 Discussion | 第77-78页 |
| Chapter Ⅴ Protein expression analysis by Indirect-ELISA | 第78-82页 |
| 5.1 Introduction | 第78页 |
| 5.2 Materials and Methods | 第78-80页 |
| 5.2.1 Preparation of antibody | 第78-79页 |
| 5.2.2 Indirect enzyme-linked immunosorbent assay(ELISA) | 第79-80页 |
| 5.3 Results | 第80-81页 |
| 5.4 Discussion | 第81-82页 |
| Chapter Ⅵ Functional analysis of myosin light chain 2 | 第82-91页 |
| 6.1 Introduction | 第82-83页 |
| 6.2 Materials and Methods | 第83-85页 |
| 6.2.1 Immunofluorescence microscopy analysis | 第83-84页 |
| 6.2.2 Full-length gene amplification,Bioinformatics and Phylogenetic analysis | 第84-85页 |
| 6.3. Results | 第85-90页 |
| 6.3.1 Sub-cellular localization analysis ofMLC2 and P7-1 in insect organs | 第85-86页 |
| 6.3.2 Sequence analysis of MLC 2 gene in S.furcifera | 第86-90页 |
| 6.4 Discussion | 第90-91页 |
| Chapter Ⅶ Discussion | 第91-95页 |
| Chapter Ⅷ Conclusion | 第95-96页 |
| References | 第96-110页 |
| Acknowledgements | 第110-112页 |
| Author biography | 第112-113页 |
| Appendix | 第113-119页 |
