| 摘要 | 第1-6页 |
| Abstract | 第6-11页 |
| Chapter 1 General Background and Analysis | 第11-37页 |
| ·Brief Introduction | 第11页 |
| ·S2P Cascades in Diverse Signaling Transductions | 第11-22页 |
| ·S2P pathway in human | 第11-13页 |
| ·RseP involved proteolytic cascade in Escherichia coli | 第13-15页 |
| ·SpoIVFB in Bacillus subtilis | 第15-17页 |
| ·RasP in Bacillus subtilis | 第17-18页 |
| ·MucP in Pseudomonas aeruginosa | 第18-19页 |
| ·MmpA in Caulobacter crescentus | 第19-20页 |
| ·HurP in Bordetella bronchiseptica | 第20页 |
| ·S2P homologs in Arabidopsis | 第20-21页 |
| ·Other newly identified S2P cascades | 第21-22页 |
| ·Variation and Conservation of S2P Cascades | 第22-34页 |
| ·Various Site-1 proteases | 第22-23页 |
| ·Conserved catalytic motif and mechanism of Site-2 proteases | 第23-31页 |
| ·Substrate recognition | 第31-34页 |
| ·Significance | 第34-35页 |
| ·Main Contents | 第35-37页 |
| Chapter 2 Slr0643 Is Essential for Acid Acclimation in Cyanobacterium Synechocystis sp. PCC 6803 | 第37-66页 |
| ·Brief Introduction | 第37-38页 |
| ·Material and Methods | 第38-42页 |
| ·Strains and culture conditions | 第38页 |
| ·Measurement of physiological parameters | 第38-39页 |
| ·Electron microscopy | 第39页 |
| ·RNA isolation | 第39-40页 |
| ·RT-qPCR validation | 第40页 |
| ·Microarray analysis | 第40-42页 |
| ·Results and Discussion | 第42-65页 |
| ·Phylogenetic analysis of S2P homologs in cyanobacteria | 第42-46页 |
| ·Characterization of Synechocystis mutant with partially disrupted slr0643 gene | 第46-51页 |
| ·Microarray analysis of wild type and slr0643 mutant under pH 7.5 and pH 6.5 | 第51-58页 |
| ·Validation of microarray data and extended acid response in selected genes | 第58-60页 |
| ·Partial disruption of slr0643 gene in Synechocystis renders acid lethality at pH 6.5 | 第60-61页 |
| ·Diminished acid inducibility in slr0643 mutant | 第61-64页 |
| ·Acid acclimation in wild type and acid lethality in slr0643 mutant | 第64-65页 |
| ·Summary | 第65-66页 |
| Chapter 3 Slr0643’Antiserum Generation, Expression in E.coli and Enzymatic Property Study | 第66-92页 |
| ·Brief Introduction | 第66-67页 |
| ·Materials and Methods | 第67-81页 |
| ·Strains and reagents | 第67-70页 |
| ·Primers used in this chapter | 第70页 |
| ·Recombinant DNA and gene expression related techniques | 第70-76页 |
| ·Western blot | 第76-78页 |
| ·His-tagged protein affinity purification | 第78-79页 |
| ·GST-tagged protein purification by affinity chromatography | 第79-81页 |
| ·Results and Discussion | 第81-91页 |
| ·Antigenic analysis of slr0643 by bioinformatic tools | 第81-82页 |
| ·Construction of expression vector pET30b+slr0643 antigenic fragment | 第82-83页 |
| ·Expression of pET30b+slr0643 antigenic fragment in E.coli strain BL21 (DE3) | 第83-84页 |
| ·Purify antigenic peptide fragment by affinity chromatography | 第84-85页 |
| ·Immune rabbit and validate the produced polyclonal antibody | 第85页 |
| ·Construction of the vectors for expressing S2P homologs in E.coli | 第85-87页 |
| ·Optimization of overexpression conditions for S2P homologs in E.coli | 第87-88页 |
| ·Characterization of the metalloprotease activity of GST fused slr0643 | 第88-91页 |
| ·Summary | 第91-92页 |
| New Findings and Innovation Points | 第92-94页 |
| Findings | 第92页 |
| Innovation Points | 第92页 |
| Perspective | 第92-94页 |
| References | 第94-105页 |
| Appendix Ⅰ | 第105-129页 |
| Appendix Ⅱ | 第129-131页 |
| 攻读硕士学位期间取得的研究成果 | 第131-132页 |
| 致谢 | 第132-133页 |
| 附件 | 第133页 |
