| Abstract | 第4-6页 |
| 摘要 | 第7-12页 |
| Chapter 1 Introduction | 第12-24页 |
| 1.1 Background and significance of this study | 第12-13页 |
| 1.2 Latest development in Synthetic Biology | 第13-15页 |
| 1.3 Biosensors and Boolean gates in synthetic biology | 第15-16页 |
| 1.4 Boolean gates based on RNAi | 第16-17页 |
| 1.5 Main and additional research content and goal | 第17-23页 |
| 1.6 Technology roadmap | 第23-24页 |
| Chapter 2 Experiment materials and methods | 第24-38页 |
| 2.1 Laboratory equipments | 第24-25页 |
| 2.2 Reagents and materials | 第25-26页 |
| 2.3 Experimental methods | 第26-38页 |
| 2.3.1 Colony picking | 第26-27页 |
| 2.3.2 Mini preparation | 第27-28页 |
| 2.3.3 Restriction digest | 第28页 |
| 2.3.4 Gel electrophoresis | 第28-29页 |
| 2.3.5 DNA elution | 第29-30页 |
| 2.3.6 Gibson Assembly (One-step isothermal DNA assembly) | 第30-31页 |
| 2.3.7 MoClo Assembly (For 0-module plasmids) | 第31-32页 |
| 2.3.8 MoClo Assembly (For 1-module plasmids) | 第32-33页 |
| 2.3.9 DNA ligation with T4 ligase | 第33-34页 |
| 2.3.10 Bacterial transformation | 第34-35页 |
| 2.3.11 Yeast transformation and integration | 第35-36页 |
| 2.3.12 Yeast cells streaking and culture | 第36-37页 |
| 2.3.13 DNA extraction from yeast cells | 第37-38页 |
| Chapter 3 DNA assembly | 第38-48页 |
| 3.1 Theory of Gibson method | 第38-41页 |
| 3.2 Theory of MoClo method | 第41-46页 |
| 3.3 Brief summary | 第46-48页 |
| Chapter 4 Constructing Boolean gates based on translational controls | 第48-61页 |
| 4.1 Construction of the target sequence that expresses fluorescence | 第48-49页 |
| 4.2 Integration of the target sequence into yeast cells | 第49页 |
| 4.3 Integration of the Dicer and the Argonaute in the yeast strain containing the target sequence (infrastructure for RNAi) | 第49-51页 |
| 4.4 Engineering RNAi with the hairpin as a siRNA precursor | 第51-52页 |
| 4.5 Engineering RNAi with convergent promoters | 第52-59页 |
| 4.5.1 Convergent promoters design with pGPD | 第52-54页 |
| 4.5.2 New convergent promoter design with pGAL1 | 第54-55页 |
| 4.5.3 Convergent promoter design with a single intron sequence | 第55-57页 |
| 4.5.4 Two different transcription units (sense-antisense design) for siRNA precursor expression | 第57-59页 |
| 4.6 Brief summary | 第59-61页 |
| Conclusion | 第61-62页 |
| References | 第62-67页 |
| Acknowledgements | 第67页 |
