| ABSTRACT IN CHINESE | 第10-12页 |
| ABSTRACT IN ENGLISH | 第12-13页 |
| 1 INTRODUCTION | 第14-22页 |
| 1.1 PATHOGENICITY OF AVIAN PATHOGENIC ESCHERICHIA COLI (APEC) | 第14-16页 |
| 1.1.1 EPIDEMIOLOGY OF AVIAN COLIBACILLOSIS | 第14页 |
| 1.1.2 VIRULENCE FACTORS OF APEC | 第14-15页 |
| 1.1.3 PREVENTION OF BROILER CHICKENS FROM APEC INFECTION | 第15-16页 |
| 1.2 LACTOBACILLUS MUCOSAL IMMUNITY PROGRESS AGAINST COUBAATLOSIS | 第16-19页 |
| 1.2.1 MECLMIISM OF MUCOSAL IMMUNITY | 第16-17页 |
| 1.2.2 LACTOBACILLUS USE AS DELIVEIY VEHICLE IN VACCINATION | 第17-18页 |
| 1.2.3 EXPRESSION STRATEGIES OF RECOMBINANT PROTEINS IN LACTIC ACID BACTERIA | 第18页 |
| 1.2.4 ADVANTAGES OF MUCOSAL IMMUNIZATION | 第18-19页 |
| 1.3 OBJECTIVE AND SIGNIFICANCE OF THE STUDY | 第19-22页 |
| 2 MATERIALS AND METHODS | 第22-39页 |
| 2.1 MATERIALS | 第22-24页 |
| 2.1.1 BACTERIAL STRAINS AND PLASMIDS | 第22页 |
| 2.1.2 PRIMERS | 第22-23页 |
| 2.1.3 EXPERIMENTAL ANIMALS | 第23页 |
| 2.1.4 CHEMICAL REAGENTS | 第23-24页 |
| 2.1.5 INSTRUMENT AND EQUIPMENT | 第24页 |
| 2.2 METHODS | 第24-39页 |
| 2.2.1 ACTIVATION OF BACTERIA | 第24页 |
| 2.2.2 E, COLI CHALLENGE TO BROILER CHICKENS | 第24-25页 |
| 2.2.3 GENOME EXTRACTION FROM APEC O78 | 第25页 |
| 2.2.4 PREPARATION OF FIMA GENE | 第25-26页 |
| 2.2.5 DNA AGAROSE GEL ELECTROPHORESIS | 第26页 |
| 2.2.6 AMPLIFICATION OF OMPC GENE | 第26-27页 |
| 2.2.7 LIGATION OF FIMA AND OMPC GENE | 第27页 |
| 2.2.8 HEAT SHOCK TRANSFORMATION OF PLASMID INTO TGI | 第27页 |
| 2.2.9 SCREENING AND IDENTIFICATION OF RECOMBINANT STRAIN | 第27-28页 |
| 2.2.10 EXTRACTION OF FIMA AND OMPC GENE FROM RECOMBINANT PLASMID | 第28-29页 |
| 2.2.11 FUSION OF FIMA AND OMPC GENE | 第29页 |
| 2.2.12 EXTRACTION OF RECOMBINANT PLASMIDS | 第29-30页 |
| 2.2.13 IDENTIFICATION OF RECOMBINANT PLASMID | 第30-31页 |
| 2.2.14 FUSION OF OMPC, FIMA AND M&DC TARGETING PEPTIDE GENES | 第31页 |
| 2.2.15 SELECTION OF LACTIC ACID BACTERIA USED FOR ANTIGEN DELIVERY CARRIER | 第31-32页 |
| 2.2.16 CONSTRUCTION OF RECOMBINANT LACTOBACILLUS EXPRESSING OMPC AND FIMA PROTEIN OF APEC | 第32-35页 |
| 2.2.17 TOLERANCE TO ACID AND BILE SALT AND GROWTH CHARACTERISTICS OF RECOMBINANT LACTOBACILLUS STRAINS | 第35-36页 |
| 2.2.18 IN VIVO COLONIZATION ABILITY OF THE RECOMBINANT LACTOBACILLUS STRAINS | 第36页 |
| 2.2.19 REGULATION OF CHICKEN BONE MARROW-DERIVED DCS (CHBM-DCS) BY RECOMBINANT LACTOBACILLUS STRAINS | 第36-37页 |
| 2.2.20 EXPRESSION AND IDENTIFICATION OF INTEREST PROTEINS | 第37页 |
| 2.2.21 IMMUNIZATION | 第37-38页 |
| 2.2.22 ELISA ANALYSIS OF ANTIBODY LEVELS | 第38页 |
| 2.2.23 CHALLENGE EXPERIMENT | 第38页 |
| 2.2.24 STATISTICAL ANALYSIS | 第38-39页 |
| 3 RESULTS | 第39-57页 |
| 3.1 CONSTRUCTION OF FUSION GENE OMPC-FIMA | 第39-42页 |
| 3.1.1 PCR AMPLIFICATION RESULT OF TARGET FIMA AND OMPC GENE | 第39页 |
| 3.1.2 IDENTIFICATION OF RECOMBINANT PMD19-T-FIMA BY ENZYME DIGESTION | 第39-40页 |
| 3.1.3 IDENTIFICATION OF PLASMID PMD19-T-FIRAA BY PCR | 第40页 |
| 3.1.4 IDENTIFICATION OF RECOMBINANT PMD19-T-OMPCBY ENZYME DIGESTION | 第40-41页 |
| 3.1.5 IDENTIFICATION OF PLASMID PMD19-T-OMPC BY PCR | 第41页 |
| 3.1.6 IDENTIFICATION OF PMD19-T-OMPC-FIMA BY ENZYME DIGESTION | 第41-42页 |
| 3.1.7 IDENTIFICATION OF PLASMID PMD19-T-OMPC-FIMA BY PCR | 第42页 |
| 3.2 IDENTIFICATION OF PLASMID PMD19-T-OMPC-FIMA-C〇I-DCPEP | 第42-44页 |
| 3.2.1 PCR IDENTIFICATION RESULT | 第42-43页 |
| 3.2.2 EN2YME DIGESTION IDENTIFICATION | 第43-44页 |
| 3.3 SELECTION OF LACTIC ACID BACTERIA FOR ANTIGEN DELIVERY CARRIER | 第44-45页 |
| 3.3.1 ADHERENCE ABILITY | 第44页 |
| 3.3.2 IMPACT ON GROWTH PERFORMANCE OF CHICKEN | 第44-45页 |
| 3.4 CONSTRUCTION OF RECOMBINANT L. SAERIMNERI (M-L 1) EXPRESSING OMPC AND FIMA PROTEIN | 第45-47页 |
| 3.5 ACID AND BILE SALT TOLERANCE AND GROWTH CHARACTER1STCS OF THE RECOMBINANT STRAINS | 第47-48页 |
| 3.6 IN VRVO COLONIZATION OF THE GASTROINTESTINAL TRACT BY RECOMBINANT LACTOBACILLUS STRAINS | 第48-50页 |
| 3.7 EXPRESSION OF THE FUSION PROTEIN | 第50页 |
| 3.8 REGULATION OF CHBM-DCS BY RECOMBINANT LACTOBACILLUS STRAINS | 第50-51页 |
| 3.9 GROWTH PROPERTIES OF CHICKEN ORALLY ADMINISTERED RECOMBINANT LACTOBACUXUS STRAINS | 第51-52页 |
| 3.10 IMMUNE RESPONSES INDUCED IN CHICKENS BY ORAL ADMINISTRATION OF THE RECOMBINANT LACTOBACILLUS STRAINS | 第52-55页 |
| 3.11 PROTECTION EFFICACY OF RECOMBINANT STRAIN AGAINST APEC | 第55-57页 |
| 4 DISCUSSIONS | 第57-62页 |
| 4.1 PHYSIOLOGICAL CHARACTERISTICS OF ISOLATED LACTOBACILLUS | 第57页 |
| 4.2 SELECTION OF PROTECTIVE ANTIGEN GENES | 第57-58页 |
| 4.3 ELECTROPORATION OF FUSION GENES INTO COMPETENT CELLS | 第58-59页 |
| 4.4 EXPRESSION OF FUSION PROTEIN IN LACTOBACILLUS | 第59页 |
| 4.5 COLONIZATION CAPACITY OF RECOMBINANT LACTOBACILLUS STRAINS | 第59-60页 |
| 4.6 IMMUNOGHNICITY IN CHICKS OF LACTOBACILLUS-BASED ORAL VACCINE | 第60-62页 |
| 5 CONCLUSIONS | 第62-63页 |
| ACKNOWLEDGMENT | 第63-64页 |
| REFERENCES | 第64-71页 |
| PAPERS PUBLISHED IN THE PERIODS OF PHD EDUCATION | 第71页 |
