| QUOTATION | 第1-4页 |
| DEDICATION | 第4-5页 |
| DECLARATION | 第5-6页 |
| CERTIFICATION | 第6-11页 |
| ABSTRACT IN ENGLISH | 第11-14页 |
| ABSTRACT IN CHINESE | 第14-16页 |
| INTRODUCTION | 第16-19页 |
| References | 第19-22页 |
| PART ONE LITERATURE REVIEW | 第22-72页 |
| CHAPTER ONE Newcastle Diseases and Iits Causative Agent | 第23-52页 |
| SECTION A Newcastle Disease Virus | 第23-34页 |
| Morphology | 第23-24页 |
| Molecualr Structure and Fuctional Characteristics of Glycoprotein | 第24-28页 |
| Interaction and Conformational Changes of Glylcoprotein during Fusion | 第28-30页 |
| Reference | 第30-34页 |
| SECTION B Newcastle Disease and Newcastle Disease Vaccines | 第34-52页 |
| Newcastle Disease | 第34-38页 |
| Research and Advances on Newcastle Disease Vaccines | 第38-45页 |
| Newcastle Disease virus "Treats" Other Disease | 第45-46页 |
| References | 第46-52页 |
| CHAPTER TWO …DNA Vaccines | 第52-72页 |
| Brief History about the Development of Vaccine | 第52-54页 |
| The Art and Principle of DNA Vaccinology | 第54-55页 |
| Research, Progress on DNA Vaccines Studies | 第55-56页 |
| Molecular Components and Construction of DNA Plasmid | 第56-57页 |
| The Generation of an Immune Response to DNA Immunization | 第57-59页 |
| Duration and Strength of the Immune Response | 第59页 |
| Route of Delivery of DNA Vaccines | 第59-60页 |
| Comparison of the Route and Influence of DNA Vaccine Delivery in Other Animal Species | 第60页 |
| DNA Delivery in Avian Species | 第60-62页 |
| Dosage and Regime | 第62-63页 |
| Immune Responses and Protection after DNA Vaccination | 第63页 |
| Advantages of DNA Vaccines, potential limitation and Future of DNA Vaccines | 第63-65页 |
| References: | 第65-72页 |
| PART TWO: EXPERIMENTAL SECTION | 第72-139页 |
| CHAPTER THREE…Cloning of Newcastle Disease Virus Hemagglutinin-Neuraminidase Gene and sequence Analysis | 第73-94页 |
| MATERIALS AND METHODS | 第73-82页 |
| Primer Design | 第73页 |
| Virus Propagation and purification (NDV F48E9 Strain) | 第73-74页 |
| Viral RNA Extraction and Synthesis of 1st Strand Cdna | 第74-75页 |
| Reverse Transcriptase Polymerase Chain Reaction | 第75-76页 |
| Construction of Recombinants plasmids pcDNA3-HN and pcDNA3-eGFP | 第76-80页 |
| Transformation of Recombinant pcDNA3-HN and pcDNA3-eGFP into DH5α | 第80页 |
| Plasmid DNA Extracton By Alkaline Lysis (Minipreparation) | 第80-82页 |
| RESULTS | 第82-90页 |
| NDV- HN Gene Amplified by RT-PCR | 第82-83页 |
| Cloning and Molecular Identification of Recombinant plasmids pcDNA3 -HN and pcDNA3 -eGFP | 第83-84页 |
| Sequencing and Sequence Analysis of NDV-HN | 第84-85页 |
| NDV-HN Nucleotide and Amino Acid Sequence | 第85-86页 |
| Deduced Amino Acids Sequences of NDV-HN Strains | 第86-89页 |
| Nucleotide Homology of NDV-HN Strains | 第89-90页 |
| Phylogenetic Tree of NDV-HN Strains | 第90页 |
| DISCUSSION | 第90-91页 |
| REFERENCES | 第91-94页 |
| CHAPTER FOUR Prokaryotic Expression of NDV-HN and eGFP in E. coli and attenuated Salmonella typhimurium | 第94-116页 |
| MATERIALS AND METHODS | 第96-106页 |
| Bacterial strains, plasmid vectors and growth media | 第96页 |
| Construction of Recombinant pGEX-6p1-Egfp | 第96-99页 |
| Preparation of Competent Cells for Heat Shock Transformation using Calcium Chloride | 第99页 |
| Heat Shock Transformation of B121(De3) Cells with pGEX-6p1-eGFP | 第99-100页 |
| Preparation of Salmonella typhimurium Competent Cells for Electroporation Transformation | 第100-102页 |
| Elecrtoporation of PGEX-6Pl-eGFP into Salmonella Typhimurium | 第102页 |
| Expression of pGEX-6P 1-eGFP IN Salmonella Host Strains | 第102-103页 |
| Fluorescent Microscopy | 第103页 |
| Construction of Recombinant pGEX-6P1-HN | 第103-105页 |
| Heat Shock Transformation of BL21(DE3) Cells with pGEX-6P1-HN Expression of pGEX-6P1-HN in E. coli BL21 | 第105-106页 |
| SDS-PAGE and Western Immunoblotting | 第106页 |
| RESULTS | 第106-110页 |
| Cloning and Molecular Identification of Recombinant Plasmid | 第106-107页 |
| Expression of eGFP in S. typhimurium and E. coli | 第107-109页 |
| Protein Analysis with SDS-PAGE for Proteins expressed in S. typhimurium | 第109-110页 |
| DISCUSSION | 第110-112页 |
| REFERENCE | 第112-116页 |
| CHAPTER FIVE …Attenuated Salmonella typhimurium as a Delivery Vehicle for Expression of pcDNA3-eGFP and NDV-HN in Hela Cells | 第116-126页 |
| MATERIALS AND METHODS | 第116-120页 |
| Bacteria and plasmids, Eukaryotic Cell Lines | 第116页 |
| Transformation of ZJ111 Competent Cells with pcDNA3- eGFP and pcDNA3- HN by elecrtoporation | 第116页 |
| Recombinant Plasmid Stability in S. typhimurium ZJ111 | 第116-117页 |
| Salmonella as a Delivery Vehicle for Expression of pcDNA3- eGFP | 第117页 |
| Cationic Liposome Mediated Transfection of HeLa Cells with pcDNA3- eGFP | 第117页 |
| Fluorescence Microscopic Examination | 第117-118页 |
| Salmonella as a Delivery Vehicle for Expression of pcDNA3- HN | 第118页 |
| Extraction of Total RNA from Cell Lysate and RT-PCR | 第118-119页 |
| SDS-PAGE and Western Immunoblotting | 第119-120页 |
| RESULTS | 第120-123页 |
| Recombinant Plasmid Stability in S. typhimurium | 第120-121页 |
| Expression of eGFP By Salmonella as a Carrier and by Cationic Liposome Mediation | 第121页 |
| RT-PCR | 第121-122页 |
| Protein Analysis with SDS-PAGE and Immunoblot Technique | 第122-123页 |
| DISCUSSION | 第123-124页 |
| REFERENCE | 第124-126页 |
| CHAPTER SIX Naked DNA Vaccination against Newcastle Disease Virus Based on Its Major Glycoprotein Genes | 第126-139页 |
| MATERIALS AND METHODS | 第126-132页 |
| Plasmid DNA Extraction:(Maxipreparation) and Purification of Plasmids | 第126-128页 |
| Vaccination of Chickens and Sampling of Blood | 第128-129页 |
| Chicken Challenge and Evaluation of Protective Efficacy | 第129页 |
| Lymphocyte Proliferation Assay (LPA) | 第129-131页 |
| ELISA for antibody titration | 第131-132页 |
| RESULTS | 第132-135页 |
| Antibody Titer Assessment | 第132页 |
| Proliferation Assay for Peripheral Blood Lymphocytes | 第132-133页 |
| Post Challenge Observation and Protection Assessment | 第133-135页 |
| DISCUSSION | 第135-137页 |
| REFERENCES | 第137-139页 |
| CONCLUSION AND RECOMMENDATION | 第139-141页 |
| ACKNOWLEDGEMENT | 第141-142页 |
