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腰带长体茧蜂嗅觉相关蛋白基因的鉴定和功能验证

摘要第6-8页
Abstract第8-9页
List of Figures第13-15页
List of Tables第15-16页
Abbreviations第16-19页
Chapter Ⅰ General Introduction第19-35页
    1.1 General overview第19页
    1.2 Olfaction in insect第19-20页
    1.3 The olfactory system第20-22页
    1.4 Olfactory related proteins第22-27页
        1.4.1 Sensillum lymph proteins第22-26页
        1.4.2 Membrane proteins第26-27页
    1.5 Activation of insect odorant receptor第27-30页
    1.6 The Macrocentrus cingulum Brischke第30-31页
    1.7 How do we know what the M. cingulum olfaction system第31-34页
        1.7.1 Olfactory protein and ligand binding第31-32页
        1.7.2 Electron microscopy第32-33页
        1.7.3 Plant odorant and behavior第33-34页
    1.8 Aims第34-35页
Chapter Ⅱ Identification of Chemosensory Receptor Genes from M. cingulum第35-49页
    2.1 Introduction第35-36页
    2.2 Materials and Methods第36-41页
        2.2.1 Insects第36页
        2.2.2 Extraction of total RNA第36-38页
        2.2.3 cDNA library construction protocol第38页
        2.2.4 Construction of cDNA library,gene identification and functional annotation第38-39页
        2.2.5 RT-PCR for expression analysis第39-41页
    2.3 Results第41-47页
        2.3.1 Antennal cDNA library of M. cingulum第41页
        2.3.2 Annotation of predicted proteins第41-42页
        2.3.3 Functional annotation by GO,COG,and KEGG第42-43页
        2.3.4 Annotation of a part of OR and IR genes第43-46页
        2.3.5 Expression patter in different tissues第46-47页
    2.4 Discussion第47-49页
Chapter Ⅲ McinOrco Identification and Expression Pattern Analysis第49-60页
    3.1 Introduction第49-50页
    3.2 Materials and Methods第50-53页
        3.2.1 Insect rearing and tissue collection第50页
        3.2.2 RNA extraction and cDNA synthesis第50页
        3.2.3 Polymerase chain reaction(PCR)第50页
        3.2.4 Cloning and sequence analysis第50-51页
        3.2.5 Expression profile of McinOrco in different tissues第51-52页
        3.2.6 Preparation of in situ hybridization probes第52页
        3.2.7 Whole mount flurescence in situ hybridization(WM-FISH)第52-53页
        3.2.8 Data analysis第53页
    3.3 Results第53-58页
        3.3.1 Cloning of Orco orthologue in M. cingulum第53-55页
        3.3.2 Tissue specific expression pattern of McinOrco第55-56页
        3.3.3 Topographic expression of mcinOrco in male and female antenna第56-58页
    3.4 Discussion第58-60页
Chapter Ⅳ Functional Characterization of McinOBP1第60-78页
    4.1 Introduction第60-62页
    4.2 Materials and Methods第62-66页
        4.2.1 Insects第62页
        4.2.2 RNA extraction and cDNA synthesis第62页
        4.2.3 Polymerase chain reaction(PCR)第62页
        4.2.4 Expression pattern of McinOBP1第62-63页
        4.2.5 Cloning and sequencing第63页
        4.2.6 Cloning of McinOBP1-wild type in expression vector第63-64页
        4.2.7 Purification of proteins第64页
        4.2.8 Preparation of site-directed mutants第64-65页
        4.2.9 Flourescence binding assays第65页
        4.2.10 Molecular modeling and ligands docking第65-66页
    4.3 Results第66-75页
        4.3.1 Cloning and sequence amalysis of McinOBP1第66页
        4.3.2 Tissues specific expression pattern of McinOBP1第66-68页
        4.3.3 Recombinant protein McinOBPl-wt expression and purification第68页
        4.3.4 Ligand protein binding specificity第68-71页
        4.3.5 Molecular modeling of McinOBP1-wt protein and ligands docking第71-73页
        4.3.6 Mutagenesis and binding assays第73-75页
    4.4 Discussion第75-78页
Chapter Ⅴ SEM and TEM of Antennal Sensilla of M. cingulum第78-93页
    5.1 Introduction第78-79页
    5.2 Materials and Methods第79-80页
        5.2.1 Insects第79页
        5.2.2 Scanning electron microscope(SEM)第79页
        5.2.3 Transmission electron microscope(TEM)第79页
        5.2.4 Terminology of sensilla第79-80页
        5.2.5 Data analysis第80页
    5.3 Results第80-89页
        5.3.1 General morphology of the antenna第80-81页
        5.3.2 Sensilla on the antenna第81-89页
            5.3.2.1 Sensilla chaetica(CHA)第81页
            5.3.2.2 Sensilla basicconica(BAS)第81-83页
            5.3.2.3 Sensilla trichodea(TRI)第83-84页
            5.3.2.4 Coeloconica sensilla(COE)第84-86页
            5.3.2.5 Sensilla placodea(PLA)第86-88页
            5.3.2.6 Sensilla campaniform(CAM)第88-89页
            5.3.2.7 The cuticular pore(CP)第89页
    5.4 Discussion第89-93页
Chapter Ⅵ Sense Organs on the Ovipositor of M. cingulum第93-104页
    6.1 Introduction第93-94页
    6.2 Materilas and Methods第94-95页
        6.2.1 Experimental insects第94页
        6.2.2 Scanning electron microscopy(SEM)第94页
        6.2.3 Transmission electron microscopy(TEM)第94-95页
    6.3 Results第95-100页
        6.3.1 Ovipositor morphology第95-97页
        6.3.2 Sensory structures on the M. cingulum ovipositor第97-100页
            6.3.2.1 Sensilla coeloconica第97-98页
            6.3.2.2 Sensilla campaniform第98页
            6.3.2.3 Sensilla trichodra第98-99页
            6.3.2.4 Secretory pores on the M. cingulum ovipositor第99-100页
    6.4 Discussion第100-104页
        6.4.1 Host selection process第100-101页
        6.4.2 Stinging mechanism第101-102页
        6.4.3 Ovipositional mechanism第102-104页
Chapter Ⅶ Behavioral Response of M.cingulum to Corn Odorants第104-112页
    7.1 Introduction第104-105页
    7.2 Materials and Methods第105-106页
        7.2.1 Insects第105页
        7.2.2 Behavioral bioassays with Y-tube olfactometer第105-106页
        7.2.3 Data analysis第106页
    7.3 Results第106-111页
    7.4 Discussion第111-112页
General Conclusion第112-116页
References第116-137页
Acknowledgement第137-139页
Resume第139-144页

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