| ABSTRACT | 第1-7页 |
| CONTENTS | 第7-11页 |
| ChapterⅠ INTRODUCTION | 第11-59页 |
| 1 -Promoter concept, activity and research development | 第12-30页 |
| ·-Structure analysis | 第16-20页 |
| ·-Promoter strength prediction and promoter cloning vectors | 第20-22页 |
| ·-Promoter of other kingdoms | 第22-25页 |
| ·-promoter research strategies and applications | 第25-30页 |
| 2 Chloramphenicol resistance | 第30-34页 |
| ·-Structure of CAT enzyme | 第31-32页 |
| ·-Inducible expression of cat gene | 第32-34页 |
| 3 Melanin | 第34-41页 |
| ·-Melanin structure | 第34-37页 |
| ·-Properties on melanin | 第37-39页 |
| ·-Tyrosinase gene cloning | 第39-40页 |
| ·-Mel system as promoter probe gene and as for expression vectors | 第40-41页 |
| 4 Pseudomonas maltophilia | 第41-45页 |
| ·-Physiological and ecological studies | 第42-44页 |
| ·-Genetic studies | 第44-45页 |
| 5 Mieroealorimetry | 第45-56页 |
| ·-instruments | 第46-48页 |
| ·-applications area | 第48-56页 |
| ·-microorganisms | 第49-54页 |
| ·-other applications | 第54-56页 |
| a -Biocompatibility | 第54页 |
| b -Ecological systems | 第54-55页 |
| c -Living materials from plants | 第55-56页 |
| 6 -The aim and significance of this study | 第56-59页 |
| ChapterⅡ promoter cloning and characterisation | 第59-81页 |
| 1 -material and methods | 第59-69页 |
| ·-material | 第59-62页 |
| ·-bacterial strains | 第59页 |
| ·-reagents | 第59-61页 |
| ·-media and components | 第61页 |
| ·-preparation of solutions | 第61-62页 |
| ·-experimental procedures | 第62-69页 |
| ·-alkaline Lysis minipreparation of plasmid DNA | 第62-63页 |
| ·-preparation of genomic DNA from bacteria | 第63页 |
| ·-purification of plasmid DNA | 第63-64页 |
| ·-agarose electrophoresis | 第64页 |
| ·-Enzymetic manipulation of DNA | 第64-65页 |
| ·-Ligation | 第65-66页 |
| ·-fresh competent cell preparation | 第66页 |
| ·-Transformation | 第66-67页 |
| ·-DNA fragments recovery | 第67页 |
| ·-Hybridization experiments | 第67-69页 |
| 2 -Results discussion | 第69-81页 |
| ·-Enzymatic digestion of pseudomonas maltophilia chromosomal DNA | 第69-70页 |
| ·Plasmid pKK232-8 extraction, purification and enzymatic digestion | 第70-72页 |
| ·Cloning of fragments containing promoter function | 第72-74页 |
| ·Ligation and transformation | 第72-74页 |
| ·Recombinants and chloramphenicol resistance | 第74-75页 |
| ·Fragments length-promoter activity relationship | 第75-77页 |
| ·Hybridization results | 第77-78页 |
| ·CAT Identification | 第78-81页 |
| ChapterⅢ Over-expression of Tyrosinase gene under the cloned promoter regulation | 第81-91页 |
| 1 method | 第81-82页 |
| ·-subcloning of mel gene into pPAS1 and construction of pAWS recombinant plasmid | 第81-82页 |
| ·-mel gene induced expression and melanin production | 第82页 |
| 2 results and discussion | 第82-91页 |
| ·-construction of pAWS and cloning results | 第82页 |
| ·-enzymatic restriction of pAWS | 第82-84页 |
| ·-melanin production by E.coli/pAWS and E.coli/pWSY | 第84-86页 |
| ·-Attempt to express Bacillus licheniformis protease gene in E.coli under control of aPAS1 promoter fragment | 第86-91页 |
| ChapterⅣ application of microcalorimetry to promoter activity | 第91-119页 |
| 1 -TG1/pPAS7 microcalorimetric study: an introduction to the gene expression calorimetric research | 第91-99页 |
| ·-material and methods | 第91-93页 |
| ·-bactrial strains | 第91页 |
| ·-calorimeter | 第91-92页 |
| ·-preparation of the sample | 第92页 |
| ·-calorimetric experiment procedure | 第92-93页 |
| ·-results | 第93-97页 |
| ·-calculation of the growth rate constant of E.coli | 第93-95页 |
| ·-inhibitory ratio and half inhibitory concentration | 第95页 |
| ·-the growth of E.coli | 第95页 |
| ·-the growth of E.coli/pKK232-8 | 第95-96页 |
| ·-the growth of E.coli/pPAS7 | 第96-97页 |
| ·-the relationship between Pm and C | 第97页 |
| ·-the relationship between tp and C | 第97页 |
| ·-discussion | 第97-99页 |
| 2 the impact of promoter activity on calorimetric information | 第99-119页 |
| ·-material and method | 第99-102页 |
| ·-preparation of samples | 第100-101页 |
| ·-calorimetric experiments procedures | 第101-102页 |
| ·-results | 第102-113页 |
| ·-discussion | 第113-119页 |
| ·-inhibitory ration and half inhibitory concentration | 第113-118页 |
| ·-promoter activity and Pm | 第118-119页 |
| CONCLUSION | 第119-127页 |
| REFERENCE | 第127-146页 |
| ACKNOWLEDGEMENT | 第146-147页 |
