| Abstract | 第8-12页 |
| 摘要 | 第13-16页 |
| ABBREVIATIONS | 第16-18页 |
| 1 INTRODUCTION | 第18-31页 |
| 1.1 Conventional castration and Its'alternative | 第18-19页 |
| 1.2 History of vaccine engineering and vaccination | 第19-23页 |
| 1.3 Salmonella enterica serovar Choleraesuis in vaccine delivery | 第23-25页 |
| 1.4 Merits of oral vaccination | 第25-26页 |
| 1.5 Kisspeptin and its ligand in reproduction contro | 第26-27页 |
| 1.6 Mechanism of Immunocastration and Kisspeptin Expression | 第27-29页 |
| 1.7 Research gap and objectives | 第29-31页 |
| 2 MATERIALS AND METHODS | 第31-50页 |
| 2.1 Materials | 第31-34页 |
| 2.1.1 Experimental Plasmid | 第31-32页 |
| 2.1.2 Main Equipment | 第32页 |
| 2.1.3 Enzymes and reagents | 第32-33页 |
| 2.1.4 Preparation of common reagents | 第33-34页 |
| 2.2 Methods | 第34-38页 |
| 2.2.1 Preparation of salmonella Electrophoretic Sensitivity | 第34-35页 |
| 2.2.2 pVAX-KISS-1-S-asd Plasmid electroporation to C500 strain | 第35页 |
| 2.2.3 Screening of Positive clones of C500(pVAX-KISS-1-S-asd) | 第35-36页 |
| 2.2.4 Extraction of bacterial genmic DNA | 第36页 |
| 2.2.5 Detection of in vitro stability of recombinant bacteria | 第36-37页 |
| 2.2.6 Preparation of C500/PKS-ASD vaccine | 第37-38页 |
| 2.3 Ethical statement and magement of experimental animals | 第38页 |
| 2.4 Vaccination of Exerimental animals | 第38-39页 |
| 2.5 Detection of anti-kisspeptin antibody concentration | 第39-42页 |
| 2.5.1 Indirect ELISA related so lution preparation | 第39-41页 |
| 2.5.2 Examination of serum testosterone level | 第41-42页 |
| 2.6 Vaccine Impacts on Libido and Testicle Morphology | 第42-43页 |
| 2.7 Histological Examination of Testis | 第43-48页 |
| 2.7.1 Testicular tissue HE staining | 第43-44页 |
| 2.7.2 Tissues messenger RNA (mRNA) level analysis | 第44-48页 |
| 2.8 Statistical analysis | 第48-50页 |
| 3 RESULTS | 第50-72页 |
| 3.1 Plasmid pKS identification site | 第50页 |
| 3.2 PCR amplification products of InvA gene | 第50-51页 |
| 3.3 Electrophoresis digestion of double endonuclease enzyme | 第51-52页 |
| 3.4 Health status of the experimental animals | 第52页 |
| 3.5 Immune responses of the rams to the oral vaccine | 第52-53页 |
| 3.6 Immunization effect on reproductive organs' tissue | 第53-57页 |
| 3.7 Seram testosterone concentration | 第57-58页 |
| 3.8 Assessment of sexual behavior | 第58页 |
| 3.9 Kisspeptin Concentration in Peripheral Blood | 第58-59页 |
| 3.10 Detection of total RNA | 第59-60页 |
| 3.11 Detecting qualities of cDNA | 第60-61页 |
| 3.12 mRNA levels in the hypothalamus | 第61-62页 |
| 3.13 LHβ, FSHβ and GnRHR RT PCR in the pituitary | 第62-63页 |
| 3.14 Expression levels of mRNA in the Pituitary | 第63-64页 |
| 3.15 LHR and FSHR RT PCR amplified to result in the testes | 第64-65页 |
| 3.16 Expression of LHR and FSHR mRNA in the Testes | 第65-66页 |
| 3.17 Extracted tissues cDNA quality testing | 第66-67页 |
| 3.18 Tissue of Digestive System KISS1 RT PCR detection | 第67-68页 |
| 3.19 Levels of KISS-1 mRNA in the other tissues | 第68-69页 |
| 3.20 GPR-54 RT PCR detected in digestive system tissues | 第69-70页 |
| 3.21 Expression levels of GPR-54 mRNA in the other tissues | 第70-71页 |
| 3.22 Influence of vaccine on growth rate | 第71-72页 |
| 4 DISCUSSION | 第72-94页 |
| 5 CONCLUSION AND RECOMMENDATION | 第94-95页 |
| REFERENCES | 第95-112页 |
| List of Publications | 第112-113页 |
| Acknowledgement | 第113页 |
