| 中文摘要 | 第6-9页 |
| ABSTRACT | 第9-11页 |
| ABBREVIATIONS | 第18-21页 |
| CHAPTER 1 RESEARCH BACKGROUND | 第21-42页 |
| 1.1 Somatic cell nuclear transfer | 第22-23页 |
| 1.2 Dolly the sheep | 第23-25页 |
| 1.3 Cloning domestic animals from somatic cells | 第25-27页 |
| 1.4 The role of MPF meiotic maturation of oocyte | 第27-29页 |
| 1.5 Enucleation of the recipient oocyte | 第29-34页 |
| 1.6 Role of histone acetylation in reprogramming of somatic nuclei following Nuclear Transfer | 第34-36页 |
| 1.7 Production of transgenic pigs | 第36-41页 |
| 1.8 Aim of this study | 第41-42页 |
| CHAPTER 2 EFFECT OF DEMECOLCINE-ASSISTED ENUCLEATION ON MPF LEVELSAND CYCLIN B1 DISTRIBUTION IN PORCINE OOCYTES | 第42-66页 |
| 2.1 Introduction | 第43-44页 |
| 2.2 Materials and Methods | 第44-54页 |
| 2.2.1 Source of reagents | 第44-45页 |
| 2.2.2 Main equipment | 第45-46页 |
| 2.2.3 Preparing Media and Materials | 第46-47页 |
| 2.2.4 Oocyte retrieval and in vitro maturation of oocytes | 第47-48页 |
| 2.2.5 Donor cell culture | 第48-49页 |
| 2.2.6 Nuclear transfer | 第49-50页 |
| 2.2.7 Embryo transfer | 第50页 |
| 2.2.8 MPF Assays | 第50-51页 |
| 2.2.9 Experimental design | 第51-53页 |
| 2.2.9.1 Experiment 1:Effects of DEM treatment on MPF levels in unenucleated MII oocytes | 第51页 |
| 2.2.9.2 Experiment 2:Effects of DEM treatment on MPF levels and the distribution of Cyclin B1 in oocytes | 第51-52页 |
| 2.2.9.3 Experiment 3:Enucleation efficiency and development of NT embryos in porcine oocytes enucleated by DEM-assisted or mechanical enucleation | 第52-53页 |
| 2.2.10 Statistical analysis | 第53-54页 |
| 2.3 Results | 第54-62页 |
| 2.3.1 Experiment 1:Effects of DEM treatment on MPF levels in MII oocytes | 第54-55页 |
| 2.3.2 Experiment 2:Effects of DEM treatment on the distribution of Cyclin B1 in oocytes | 第55-58页 |
| 2.3.3 Experiment 3:Enucleation efficiency and development of NT embryos in porcine oocytes by DEM-assisted enucleation or mechanical enucleation | 第58-62页 |
| 2.4 Discussion | 第62-66页 |
| CHAPTER 3 EFFECT OF CUDC-101 TREATMENT AFTER SOMATIC CELL NUCLEA | 第66-74页 |
| 3.1 Introduction | 第67-68页 |
| 3.2 Materials and methods | 第68-69页 |
| 3.2.1 In vitro-matured oocytes | 第68页 |
| 3.2.2 Isolation and culture of pig somatic cells | 第68页 |
| 3.2.3 Nuclear transfer | 第68页 |
| 3.2.4 Transfer of embryos and pregnancy determination | 第68-69页 |
| 3.2.5 Experimental design | 第69页 |
| 3.2.6 Statistical analysis | 第69页 |
| 3.3 Results | 第69-72页 |
| 3.4 Discussion | 第72-74页 |
| CHAPTER 4 PRODUCTION OF TRANSGENIC MINIATURE PIGLETS BY TRANSFEROF IFN-γ,SCFV AND IS GENE TRANSFECTED SOMATIC CELLS INTOENUCLEATED OOCYTES | 第74-87页 |
| 4.1 Introduction | 第75-76页 |
| 4.2 Materials and Methods | 第76-80页 |
| 4.2.1 Plasmids,chemicals and animals | 第76-78页 |
| 4.2.2 Donor cell cuture | 第78页 |
| 4.2.3 Transfection of resistance gene into ear fibroblasts | 第78页 |
| 4.2.4 Retrieval and vitro maturation of oocytes | 第78页 |
| 4.2.5 Nuclear transfer | 第78页 |
| 4.2.6 Embryo transfer | 第78-79页 |
| 4.2.7 Quality evaluation of embryos cultured in vitro | 第79页 |
| 4.2.8 Identification of positive clones | 第79-80页 |
| 4.2.9 Statistical analysis | 第80页 |
| 4.3 Results | 第80-84页 |
| 4.3.1 Embryo development following SCNT with nuclei obtained from IFN-γ transfected or non-transfected fibroblasts | 第80页 |
| 4.3.2 Generation of transgenic Wuzhishan miniature pigs | 第80-84页 |
| 4.4 Discussion | 第84-87页 |
| ACKNOWLEDGMENT | 第87-90页 |
| REFERENCES | 第90-105页 |
| PUBLICATION | 第105-107页 |
