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Cloning,Expression,and Functional Analysis of two Acetylcholinesterase Genes in Spodoptera Litura

Abstract第7-9页
摘要第10-12页
Chapter 1. LITERATURE REVIEW第12-41页
    1.1 Introduction第12-18页
        1.1.1 Lepidopteran insect pests第12-14页
        1.1.2 Common cutworm, Spodoptera litura Fabricius: Insect Pest第14页
        1.1.3 Common Name第14-15页
        1.1.4 Classification第15页
        1.1.5 Economic importance第15页
        1.1.6 Morphology第15页
        1.1.7 Life cycle第15-16页
        1.1.8 Insecticide Resistance in Spodoptera litura第16-17页
        1.1.9 Plant-Insect Interactions第17-18页
    1.2 Acetylcholinesterase第18-25页
        1.2.1 Distribution of Acetylcholinesterase第18-19页
        1.2.2 Insecticide Target Sites and Structural Biology of Acetylcholinesterase第19-21页
        1.2.3 Acetylcholinesterase insecticide Resistance Mechanisms第21-24页
        1.2.4 Gene encoding acetylcholinesterase in insects第24-25页
    1.3 RNA interference第25-31页
        1.3.1 RNA interference第25页
        1.3.2 A short history of the innovation of RNAi第25-27页
        1.3.3 RNAi Pathways and Mechanism第27-28页
        1.3.4 Use of RNAi第28-31页
    1.4 RNAi in insects第31-40页
        1.4.1 Preliminary RNAi trials in insects第31-32页
        1.4.2 Study on the function of insect genes using RNAi methods第32-35页
        1.4.3 Techniques of dsRNA transfer to Insects第35-36页
        1.4.4 Use of RNAi in insect pest control strategies第36-39页
        1.4.5 Susceptibility to RNAi第39-40页
    1.5 Objectives of the study第40-41页
Chapter 2. Cloning and expression of two Ace genes in Spodoptera litura第41-72页
    2.1 Introduction第41-42页
    2.2 Materials and Methods第42-45页
        2.2.1 Laboratory Equipment:第42-43页
        2.2.2 Reagents and Expression vectors第43-44页
        2.2.3 Other Reagents第44-45页
    2.3 Methods第45-56页
        2.3.1 Cloning of cDNA fragments of two SlAce genes第45页
        2.3.2 Insects第45页
        2.3.3 RNA extraction第45-46页
        2.3.4 First Strand cDNA Synthesis第46页
        2.3.5 Polymerase Chain Reaction (PCR)第46-47页
        2.3.6 Purification of DNA using DNA Gel Extraction Kit第47-49页
        2.3.7 DNA ligation with pGEM-T Easy Vector第49页
        2.3.8 Transformation of plasmid DNA into Bacteria host (E. coli)第49-51页
        2.3.9 Bacteria PCR第51页
        2.3.10 Phylogenetic analysis第51-53页
        2.3.11 SlAce genes expression pattern in different developmental stages and tissues第53-56页
        2.3.12 Statistical analysis第56页
    2.4 Results第56-68页
        2.4.1 Sequence analysis of Spodoptera litura AChE genes第56-66页
        2.4.2 SlAce genes expression profiles in S. litura第66-68页
    2.5 Discussion第68-70页
    2.6 Conclusion第70-72页
Chapter 3. Functional analysis of two Ace genes in Spdoptera litura第72-83页
    3.1 Introduction第72-73页
    3.2 Methods第73-75页
        3.2.1 dsRNA synthesis and application第73-74页
        3.2.2 SlAce knockdown determination using qRT-PCR第74页
        3.2.3 Statistical analysis第74-75页
    3.3 Results第75-81页
        3.3.1 Gene silencing through larvae injection of dsSlAce第75-76页
        3.3.2 Injection with dsRNA delays pupation and emergence第76-77页
        3.3.3 dsAce2 injection affects egg laying, egg hatching and progeny development第77-81页
    3.4 Discussion第81-83页
References第83-102页
Appendix I第102-104页
Appendix II第104-105页
Scientific Publications第105-106页
Acknowledgements第106-108页

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