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拟南芥组蛋白甲基转移酶SDG25和组蛋白分子伴侣NAP1的功能研究

摘要第7-8页
Abstract第8页
INTRODUCTION第9-60页
    Ⅰ Chromatin organization and remodeling第10-35页
        Ⅰ-1 Euchromatin and Heterochromatin第11-12页
        Ⅰ-2 Epigenetic regulation and chromatin structure第12页
        Ⅰ-3 ATP-dependent chromatin remodeling第12-13页
        Ⅰ-4 Histone va riants第13-15页
            Ⅰ-4-1 H2A variants第14-15页
            Ⅰ-4-2 H3 variants第15页
        Ⅰ-5 Histone covalent modifications第15-22页
            Ⅰ-5-1 Histone Acetylation and deacetylation第16页
            Ⅰ-5-2 Histone Ubiquitination第16-17页
            Ⅰ-5-3 Histone Phsophorylation第17-18页
            Ⅰ-5-4 Histone Methylation第18-22页
        Ⅰ-6 Nucleosome assembly and Histone chaperones第22-35页
            Ⅰ-6-1 CAF-1(Chromatin Assembly Factor-1)第24-27页
            Ⅰ-6-2 HIRA(Histone regulatory homolog A)第27-28页
            Ⅰ-6-3 Asf1(Anti-silencing function 1)第28-29页
            Ⅰ-6-4 FACT(Faciliates chromatin transcription)complex第29-30页
            Ⅰ-6-5 NAP1(Nucleosome Assembly Protein 1)第30-35页
                Ⅰ-6-5-1 Histone binding and Chaperone activity of NAP1第32-33页
                Ⅰ-6-5-2 TAF-1β/SET第33-34页
                Ⅰ-6-5-3 Plant homoOgue of NAP1 family genes第34-35页
    Ⅱ. DNA damage and repair第35-50页
        Ⅱ-1. Stress induced DNA damage第35-36页
        Ⅱ-2. DNA damage repair pathways第36-37页
        Ⅱ-3. Photoreactivation repair第37-38页
        Ⅱ-4. Nucleotide excision repair第38-42页
        Ⅱ-5. DNA double-strand break repair第42-50页
            Ⅱ-5-1. Non-homologous end joining第44-46页
            Ⅱ-5-2. Homologous recombination第46-50页
    Ⅲ DNA repair in the context of chromatin第50-58页
        Ⅲ-1 Histone modifications in DNA damage response第51-55页
            Ⅲ-1-1 H2A/H2AX phosphorylation in DNA damage respose第52-53页
            Ⅲ-1-2 Histone H2A/H2B ubiquitination in DNA Repair第53页
            Ⅲ-1-3 Histone acetylation in DNA Repair第53-54页
            Ⅲ-1-4 histone H3/H4 methylation in DNA Repair第54-55页
        Ⅲ-2 Chromatin remodeling in DNA Repair第55-56页
        Ⅲ-3 Knowledge in plants第56-58页
    Ⅳ Thesis work第58-60页
RESULTS第60-111页
    Ⅰ SDG25 Encodes a Histone Methyltransferase and is Involved in FLCActivation and Repression of Flowering第61-72页
    Ⅱ Functional analysis of Arabidopsis Nucleosome Assembly Protein1(NAP1)第72-111页
        Ⅱ-1 Molecular and reverse genetic characterization of Nucleosome Assembly Protein1(NAP1)genes unravels their function in transcription and nucleotide excision repair in Arabidopsis thaliana第73-86页
        Ⅱ-2 A Truncated Arabidopsis Nucleosome Assembly Proteinl,AtNAP1;3T, Alters Plant Growth Responses to Abscisic Acid and Salt in the Atnap1;3-2 Mutant第86-99页
        Ⅱ-3 NAP1 family histone-chaperones are required for somatic homologous recombination in Arabidopsis第99-111页
GENERAL DISCUSSION AND PERSPECTIVES第111-118页
    Ⅰ General Discussion第112-117页
        Ⅰ-1.Roles of Histone methylation in transcription activation and flowering time control第112-114页
        Ⅰ-2.Roles of histone chaperones in regulation of plant development and DNA Repair第114-117页
    Ⅱ Perspectives第117-118页
MATERIAL AND METHODS第118-136页
    Ⅰ materials第119-126页
        Ⅰ-1 plant materials第119页
        Ⅰ-2 Bacteria strains第119-120页
            Ⅰ-2-1 Escherichia coli第119-120页
            Ⅰ-2-2 Agrobacterium tumefaciens第120页
        Ⅰ-3 Cloning and expression vectors第120-122页
        Ⅰ-4 Antibodies第122-123页
        Ⅰ-5 Antibiotics第123页
        Ⅰ-6 Chemicals第123-124页
        Ⅰ-7 Oligos第124-126页
    Ⅱ methods第126-136页
        Ⅱ-1 General techniques in molecular biology第126-132页
            Ⅱ-1-1 Vector construction第126-128页
                Ⅱ-1-1-1 PCR(Polymerase Chain Reaction)第126页
                Ⅱ-1-1-2 Restriction enzyme digestion第126页
                Ⅱ-1-1-3 Agarose gel electrophoresis第126页
                Ⅱ-1-1-4 Ligation第126页
                Ⅱ-1-1-5 Competent cell preparation第126-127页
                Ⅱ-1-1-6 Transformation of competent cells第127-128页
                Ⅱ-1-1-7 Mini-preparation of plasmid DNA by alkaline lysis with SDS第128页
            Ⅱ-1-2 Genomic DNA isolation第128-129页
            Ⅱ-1-3 RNA isolation and analysis第129-130页
                Ⅱ-1-3-1 Total RNA extraction第129页
                Ⅱ-1-3-2 Reverse transcription第129-130页
            Ⅱ-1-4 Quantitative PCR analysis第130页
            Ⅱ-1-5 Protein expression,purification and detection第130-132页
                Ⅱ-1-5-1 Protein expression in E.Coli第130页
                Ⅱ-1-5-2 Purification of expressed proteins from E.Coli第130-131页
                Ⅱ-1-5-3 Separation of proteins by SDS-PAGE第131页
                Ⅱ-1-5-4 Western blot第131-132页
                Ⅱ-1-5-5 In vitro histone methytranferase activity assay第132页
        Ⅱ-2 Tobacco BY2 cells culture and transformation第132-133页
            Ⅱ-2-1 TBY2 cell culture第132页
            Ⅱ-2-2 TBY2 cell transformation第132-133页
        Ⅱ-3 Arabidopsis thaliana culture and transformation第133页
            Ⅱ-3-1 Arabidopsis culture第133页
            Ⅱ-3-2 Arabidopsis transformation by flower dip method第133页
        Ⅱ-4 Phenotype analysis of ABA and salt treatment第133-134页
        Ⅱ-5 UV treatment and In Vitro DNA Repair Assay第134页
        Ⅱ-6 The GUS-based HR reporter lines analysis第134-136页
            Ⅱ-6-1 Growth conditions and stress treatments第135页
            Ⅱ-6-2 Histochemical staining and HR assessment第135-136页
REFERENCE第136-156页
ABBREVIATIONS第156-157页
ACKNOWLEDGEMENTS第157-158页

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