| Abstract | 第1-5页 |
| 中文摘要 | 第5-11页 |
| Chapter 1: Litterature review | 第11-23页 |
| ·The Mpk1 protein | 第11-12页 |
| ·Pheromone-induced Mpk1 activation | 第12-13页 |
| ·The Bni1p, Pea2p and Spa2p proteins | 第13-19页 |
| ·Genes Required for Normal Pheromone-Induced Cell polarization in Saccharomyces cerevisiae | 第13-14页 |
| ·Polarized Growth Controls Cell Shape and Bipolar Bud Site Selection in Saccharomyces cerevisiae | 第14-16页 |
| ·Interaction of Spa2p with cell polarity proteins and signaling components involved in cell morphogenesis | 第16-17页 |
| ·Spa2p: a scaffold-like protein | 第17-18页 |
| ·Role of scaffold molecules in MAP Kinase signaling | 第18-19页 |
| ·The cell wall integrity pathway of Saccharomyces Cereviciea | 第19-23页 |
| ·Targets of the MAP Kinase pathway | 第20-21页 |
| ·MKK1 and MKK2 function | 第21-23页 |
| Chapter 2: Material and methods | 第23-45页 |
| ·Material | 第23-31页 |
| ·Plasmid, primers and strains | 第23-25页 |
| ·Plasmid | 第23页 |
| ·Primers | 第23-24页 |
| ·Strains | 第24-25页 |
| ·Equipments used in this work | 第25-26页 |
| ·Reagents used in this work | 第26-27页 |
| ·Culture medium | 第27-28页 |
| ·The main solution used in this work | 第28-31页 |
| ·Methodology | 第31-45页 |
| ·E.coli top10 preparation & transformation | 第31-32页 |
| ·CaCl_2 method | 第31-32页 |
| ·Plasmid extraction from E.coli in small scale (CTAB method) | 第32页 |
| ·DNA digestion by restriction endonuclease | 第32-33页 |
| ·DNA agarose gel electrophoresis | 第33-34页 |
| ·Purification of DNA from agarose gel | 第34页 |
| ·DNA ligation | 第34-35页 |
| ·PCR (Polymerase chain reaction) | 第35-36页 |
| ·Yeast transformation | 第36-37页 |
| ·Fast method of yeast transformation | 第36页 |
| ·High efficient transformation | 第36-37页 |
| ·Plasmid extraction from yeast cells | 第37-38页 |
| ·Mating and sporulation of differents yeast mating type | 第38页 |
| ·Tetrad dissection | 第38页 |
| ·confirmation of yeast mating type | 第38-39页 |
| ·Gene deletion by one-step replacement | 第39-40页 |
| ·Quantitative mating assay | 第40页 |
| ·Samples preparation for western blotting | 第40-42页 |
| ·Growth conditions | 第40页 |
| ·Cell lysis | 第40-41页 |
| ·Protein determination (Bio-Rad method) | 第41-42页 |
| ·Western blotting | 第42-45页 |
| ·SDS-PAGE electrophoresis | 第42页 |
| ·Setting up the western | 第42-43页 |
| ·Taking down the membrane | 第43页 |
| ·Primary antibody | 第43页 |
| ·Secondary antibody binding | 第43-44页 |
| ·Reaction with chemiluminescent reagent and Development of the film | 第44-45页 |
| Chapter 3: Results and discussions | 第45-57页 |
| ·Construction of bni1Δand spa2△ strains | 第45-48页 |
| ·Construction of spa2△ strain | 第45-47页 |
| ·Construction of bni1Δ mutation strain | 第47-48页 |
| ·Making bni1Δ mkk2Δ, spa2Δ mkk2Δ and pea2Δ mkk2Δ double mutants | 第48-51页 |
| ·Mating type determination by PCR | 第51-52页 |
| ·Mating efficiency between the mating types of each strain | 第52-55页 |
| ·Mpk1p activation upon mating pheromone induction | 第55-57页 |
| Chapter 4: Conclusion and perspectives | 第57-59页 |
| ·General conclusion | 第57-58页 |
| ·Perspectives | 第58-59页 |
| References | 第59-65页 |
| Abreviations | 第65-66页 |
| Acknowledgments | 第66页 |
