| 摘要 | 第5-7页 |
| ABSTRACT | 第7-9页 |
| Chapter 1 Review of Literature | 第12-20页 |
| 1.1 Plants and insects interactions | 第12-13页 |
| 1.1.1 Defense responses in plants | 第12-13页 |
| 1.1.2 Adaption of insects to plant defense compounds | 第13页 |
| 1.2 Introduction to P450 monooxygenases | 第13-17页 |
| 1.2.1 Endogenous functions of P4505 | 第14页 |
| 1.2.2 Role of P4505 in xenobiotic metabolism | 第14-15页 |
| 1.2.3 Induction of P450 genes by different xenobiotic compounds | 第15-17页 |
| 1.3 RNA interference | 第17-18页 |
| 1.4 Objectives and significance | 第18-20页 |
| Chapter 2 Functional characterization of cis-acting elements mediating flavoneinducible expression of CYP321A1 | 第20-45页 |
| 2.1 Introduction | 第20-21页 |
| 2.2 Materials and methods | 第21-30页 |
| 2.2.1.Cell culture | 第21页 |
| 2.2.2.Analysis of CYP321A1 expression in H. zea fatbody cells | 第21-24页 |
| 2.2.3 Construction of CYP321A1 promoter-pGL3 constructs | 第24-27页 |
| 2.2.4 Transient transfection and dual luciferase assay | 第27-28页 |
| 2.2.5 Electrophoretic gel mobility shift assay | 第28-30页 |
| 2.3 Results | 第30-41页 |
| 2.3.1 CYP321A1 transcription in H. zea fatbody cell | 第30-31页 |
| 2.3.2.Putative cis elements within the CYP321A1 promoter and 5’untranslated region | 第31页 |
| 2.3.3 Roles of 5’UTR in the transcriptional regulation of CYP321A1 | 第31-33页 |
| 2.3.4 Roles of H2151-3 in the transcriptional regulation of CYP321A1 | 第33-34页 |
| 2.3.5 Localization of cis-acting elements by progressive 5’deletions | 第34-35页 |
| 2.3.6 Fine mapping of XRE-Fla, the CYP321A1 essential cis element | 第35-39页 |
| 2.3.7 Mutational analysis of XRE-Fla | 第39页 |
| 2.3.8 H. zea fatbody cell nuclear proteins bind to the XRE-Fla | 第39-41页 |
| 2.4 Discussion | 第41-45页 |
| Chapter 3 Xanthotoxin-induced expression of CYP321A1 gene is mediated by ARE | 第45-59页 |
| 3.1 Introduction | 第45-47页 |
| 3.2 Materials and methods | 第47-48页 |
| 3.2.1 Cell culture | 第47页 |
| 3.2.2 Total RNA preparation and RT-PCR | 第47页 |
| 3.2.3 Construction of CYP321A1 promoter-pGL3 constructs | 第47页 |
| 3.2.4 Transient transfection and dual luciferase assay | 第47页 |
| 3.2.5 Induction of CYP321A1 by mixtures of xanthotoxin plus flavone | 第47-48页 |
| 3.3 Results | 第48-55页 |
| 3.3.1 Xanthotoxin induces CYP321A1 transcription | 第48页 |
| 3.3.2 Roles of 5’UTR in xanthotoxin-induced expression of CYP321A1 | 第48-51页 |
| 3.3.3 Roles of HzIs1-3 in the transcriptional regulation of CYP321A1 | 第51页 |
| 3.3.4 Localization of cis-acting elements by progressive 5’deletions | 第51页 |
| 3.3.5 Fine mapping of the CYP321A1 essential xenobiotic response element to xanthotoxin | 第51-54页 |
| 3.3.6 Mutational analysis of xenobiotic response element to xanthotoxin | 第54页 |
| 3.3.7 Induction of CYP321A1 by mixture of xanthotoxin plus flavone | 第54-55页 |
| 3.4 Discussion | 第55-59页 |
| Chapter 4 Chapter 4 ARE/CncC mediates CYP321A1 gene expression in response to plant allelochemicals | 第59-76页 |
| 4.1 Introduction | 第59-61页 |
| 4.2 Materials and methods | 第61-66页 |
| 4.2.1.Cell culture | 第61页 |
| 4.2.2 Construction of plasmids | 第61-62页 |
| 4.2.3 Transient transfection and dual luciferase assay | 第62页 |
| 4.2.4 Coexpression of dCncC protein with the wild-type CYP321A1 promoter. | 第62-63页 |
| 4.2.5 Synthesis of dsRNAs and hCncC RNA interference assay | 第63页 |
| 4.2.6 Total RNA preparation and RT-PCR | 第63-65页 |
| 4.2.7 Measurement of ROS production | 第65页 |
| 4.2.8 Electrophoretic gel mobility shift assay (EMSA) | 第65-66页 |
| 4.3 Results | 第66-74页 |
| 4.3.1 DEM and tBHQ induced transcription of CYP321A1 promoter | 第66-67页 |
| 4.3.2 Inhibition effect of blocker of NADPH oxidase (Nox) enzymes (DPI) | 第67-69页 |
| 4.3.3 Coexpression of Drosophila CncC protein with the wild-type CYP321A1 promoter | 第69-71页 |
| 4.3.4 DsRNA-Mediated inhibition of CYP321A1 promoter activity in the H. zea fatbody cells | 第71-72页 |
| 4.3.5 EMSA | 第72-74页 |
| 4.4 Discussion | 第74-76页 |
| Chapter 5 Conclusions | 第76-77页 |
| References | 第77-88页 |
| Acknowledgments | 第88-89页 |
| Curriculum Vita | 第89页 |
