| Abstract | 第1-14页 |
| Abstract (in Chinese) | 第14-16页 |
| Introduction | 第16-21页 |
| Chapter 1-Rabbit hemorrhagic disease and its causative agent | 第21-34页 |
| Section A-Rabbit hemorrhagic disease: An introduction | 第21-29页 |
| Section B-Rabbit hemorrhagic disease virus: structure, genome and variants | 第29-34页 |
| Chapter 2-VP60 capsid protein: Structure and expression as subunit vaccine in heterologous systems | 第34-50页 |
| ·RHDV capsid structure | 第34-35页 |
| ·Expression of VP60 protein as subunit vaccine in heterologous systems | 第35-42页 |
| References | 第42-50页 |
| Chapter 3-Cloning of RHDV VP60 capsid gene fragments | 第50-72页 |
| ·Design of primers | 第50-51页 |
| ·Isolation of total RNA from RHDV-infected livers | 第51-52页 |
| ·Reverse transcriptase-polymerase chain reaction (RT-PCR) | 第52-55页 |
| ·Cloning of VP60 gene fragments into pUC19 plasmid | 第55-64页 |
| ·Preparation of inserts | 第56-57页 |
| ·Preparation of the pUC19 plasimd for ligation | 第57-60页 |
| ·Ligation of insert DNAs with pUC19 plasmid DNA | 第60-61页 |
| ·Transformation of competent E.coli DH5α cells with pUC constructs | 第61-64页 |
| ·Results | 第64-70页 |
| ·Discussion | 第70-72页 |
| Chapter 4-Expression of VP60 gene fragments in E.coli | 第72-100页 |
| ·Overview of pET Expression System | 第72-74页 |
| ·Cloning of VP60 gene fragments into pET-30a | 第74-85页 |
| ·Preparation of VP60 gene inserts | 第74-79页 |
| ·Preparation of the pET30-a plasimd for ligation | 第79-81页 |
| ·Ligation of insert DNAs with pET-30a plasmid DNA | 第81-82页 |
| ·Transformation of competent E.coli cells with pET constructs | 第82-85页 |
| ·Expression of full and partial-length RHDV VP60 Proteins in E.coli: induction, identification and purification | 第85-95页 |
| ·Optimization of expression conditions | 第85-87页 |
| ·Identification of proteins by SDS-polyacrylamide gel electrophoresis | 第87-91页 |
| ·Expression and purification of His-tagged proteins | 第91-95页 |
| ·Regeneration of Ni-NTA agarose beads | 第95页 |
| ·Results | 第95-98页 |
| ·Discussion | 第98-100页 |
| Chapter 5-Immunoreactivity of full and partial-length RHDV VP60 Proteins | 第100-115页 |
| ·Raising RHDV anti-serum | 第101-102页 |
| ·Determination of protein concentrations by the method of Bradford | 第102-104页 |
| ·Testing the immunoreactivity of VP60 proteins and development of an ELISA formeasuring antibody response to vaccination | 第104-109页 |
| ·Enzyme-linked immunosorbent assay (ELISA) | 第104-106页 |
| ·Western blotting | 第106-109页 |
| ·Evaluation of ELISA using the field serum samples | 第109-110页 |
| ·Results | 第110-113页 |
| ·Discussion | 第113-115页 |
| Chapter 6-Expression of VP60 gene fragments in Sf9 insect cell line | 第115-138页 |
| ·Overview of Bac-to-Bac baculovirus expression system | 第116-118页 |
| ·Cloning of VP60 gene fragments into pFastBac HT A plasmid | 第118-124页 |
| ·Preparation of inserts | 第118-121页 |
| ·Preparation of the pFastBac HT A plasimd for ligation | 第121-122页 |
| ·Ligation of insert DNAs with pFastBac HT A plasmid DNA | 第122页 |
| ·Transformation of competent E.coli cells with pFastBac constructs | 第122-124页 |
| ·Generation of recombinant baculoviruses and protein expression | 第124-129页 |
| ·Generation the recombinant bacmids | 第124-126页 |
| ·Production of recombinant baculoviruses | 第126-129页 |
| ·Expressing the recombinant proteins | 第129页 |
| ·Testing the immunoreactivity of VP60 proteins | 第129-130页 |
| ·Western blot analysis | 第130页 |
| ·Immunofluorescence assay | 第130页 |
| ·VLPs preparation and electron microscopy | 第130-131页 |
| ·Results | 第131-136页 |
| ·Discussion | 第136-138页 |
| Conclusion | 第138-139页 |
| References | 第139-140页 |
