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腰带长体茧蜂化学感受基因的克隆、表达及功能研究

Abstract (In Chinese)第6-7页
Abstract (In English)第7-8页
List of Abbreviations第13-15页
Chapter Ⅰ General Introduction第15-33页
    1.1 Olfaction第15-16页
    1.2 Insect olfaction第16页
    1.3 Molecular basis of insect olfaction第16-18页
    1.4 Peripheral events in insect olfaction第18-26页
        1.4.1 Odorant binding protein (OBP)第19-25页
            1.4.1.1 Classification of OBPs第19-20页
            1.4.1.2 Insect OBPs structure第20-23页
            1.4.1.3 Functions of OBPs第23-25页
        1.4.2 Membrane Protein第25-26页
    1.5 Odor coding and olfactory signal transduction第26-29页
    1.6 Odor processing in sensory organs第29页
    1.7 The Macrocentrus cingulm Brischke第29-30页
    1.8 M. cingulum olfaction system and odorant protein binding assay第30-32页
    1.9 Research significance and objectives第32-33页
Chapter Ⅱ: Gene set of Olfactory Proteins in the Polyembryonic Endoparasitoid Macrocentrus cingulum第33-68页
    2.1 Introduction第33-35页
    2.2 Material and Methods第35-46页
        2.2.1 Insects第35-36页
        2.2.2 Extraction of total RNA第36-37页
        2.2.3 Antennal transcriptome generations第37页
        2.2.4 Gene identification and annotation第37-38页
        2.2.5 Phylogenetic analysis第38页
        2.2.6 RT-PCR analysis第38-44页
        2.2.7 Quantitative real-time PCR measurement第44-46页
    2.3 Results第46-63页
        2.3.1 Sequencing and assembly第46页
        2.3.2 Functional Annotation第46-50页
        2.3.3 Identification of olfactory gene families第50-61页
            2.3.3.1 Odorant binding proteins第50页
            2.3.3.2 Chemosensory proteins第50-52页
            2.3.3.3 Odorant receptors第52-58页
            2.3.3.4 Gustatory receptors第58-59页
            2.3.3.5 Ionotropic receptors第59-61页
        2.3.4 Tissue and sex specific expression profile of candidate Macrocentrus cingulum chemosensory receptors第61-63页
    2.4 Discussion第63-68页
Chapter Ⅲ: Thermal Stability and Binding Affinity of Two Odorant Binding Proteins in Macrocentrus cingulum第68-83页
    3.1 Introduction第68-69页
    3.2 Materials and Methods第69-74页
        3.2.1 Insects culture第69-70页
        3.2.2 Total RNA isolation and cDNA synthesis第70页
        3.2.3 Polymerase chain reaction (PCR)第70页
        3.2.4 Cloning and sequencing第70-71页
        3.2.5 Expression pattern with RTq-PCR第71页
        3.2.6 Cloning of McinOBP-wild types in expression vectors第71页
        3.2.7 Purification of proteins第71-72页
        3.2.8 Preparation of site-directed mutants第72页
        3.2.9 Selection of corn odorants第72-73页
        3.2.10. Circular dichroism (CD) measurements第73页
        3.2.11. Thermal stability measurements with CD第73-74页
    3.3 Results第74-81页
        3.3.1 Cloning and sequence analysis of McinOBP2 and McinOBP3第74页
        3.3.2 Tissues expression of McinOBP2 and McinOBP3 in M. cingulum第74-75页
        3.3.3 Secondary structure content prediction第75-76页
        3.3.4 Thermal stability of wild type and mutant type第76-77页
        3.3.5 Odorant affinity with wild type and mutant第77-81页
    3.4 Discussion第81-83页
Chapter Ⅳ: C-terminus a Methionene Specifically Involved in Binding Corn Odorants to McinOBP4第83-96页
    4.1 Introduction第83-84页
    4.2 Materials and Methods第84-88页
        4.2.1 Sample collection and RNA extraction第84-85页
        4.2.2 Polymerase chain reaction (PCR)第85页
        4.2.3 Cloning and sequencing第85页
        4.2.4 McinOBP4 expression and purification第85-86页
        4.2.5 Preparation of mutants第86页
        4.2.6 Distribution pattern of McinOBP4第86-87页
        4.2.7 Fluorescence measurements第87页
        4.2.8 Competitive binding assay第87页
        4.2.9 Dissociation constants (K_D) calculation第87页
        4.2.10 Molecular modelling第87-88页
    4.3 Results第88-93页
        4.3.1 Characteristics of McinOBP4 protein第88-89页
        4.3.2 Tissue distribution specificity of McinOBP4第89页
        4.3.3 Ligand-binding of recombinant McinOBP4-Wt第89-91页
        4.3.4 Mutant binding assay第91-92页
        4.3.5 Molecular modeling第92-93页
    4.4 Discussion第93-96页
General Conclusion第96-98页
References第98-120页
Acknowledgement第120-122页
Resume第122-128页

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