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粗糙脉孢菌异染色质基因met-8调控机制研究

摘要第3-4页
ABSTRACT第4-5页
ABBREVATIONS第6-10页
CHAPTER 1 INTRODUCTION第10-32页
    1.1 Chromatin第10-26页
        1.1.1 Euchromatin and heterochromatin第11-12页
        1.1.2 Discovery of heterochromatin第12页
        1.1.3 Heterochromatin architecture第12-13页
        1.1.4 Heterochromatin structure第13-15页
        1.1.5 Small RNAsare involvedin the formation of heterochromatin第15-16页
        1.1.6 Maintenance of heterochromatin第16-17页
        1.1.7 Heterochromatin barrier elements第17-18页
        1.1.8 Nature of heterochromatin第18-19页
        1.1.9 Heterochromatic environments affect gene expression第19-22页
        1.1.10 Histone modification第22-26页
        1.1.11 Chromatin remodeling factors第26页
    1.2 Regulation of gene transcription第26-29页
        1.2.1 Transcription factors第26-28页
        1.2.2 Promoter第28-29页
    1.3 Identification of heterochromatin gene met-8第29-30页
        1.3.1 Methionine第29-30页
        1.3.2 Addition of methionine represses the MET regulon第30页
    1.4 Objectives of this study第30-32页
CHAPTER 2 MATERIALS AND METHODS第32-57页
    2.1 Media and antibody第32-34页
        2.1.1 Trace element stock and solution第32页
        2.1.2 50×Vogel's salt第32页
        2.1.3 Minimal media(0.1% Glucose)第32页
        2.1.4 Minimal media(2% Glucose)第32页
        2.1.5 10×Fig's第32-33页
        2.1.6 Bottom agar/Top agar第33页
        2.1.7 0.5 MQA第33页
        2.1.8 Minimal slants第33页
        2.1.9 Race tube medium第33页
        2.1.10 LB liquid medium第33-34页
        2.1.11 LB solid medium第34页
        2.1.12 Antibody第34页
    2.2 E.coli strains第34页
    2.3 Mutant vector第34-40页
    2.4 Inoculated strains method第40-41页
    2.5 Typical protocol for making construct第41-42页
    2.6 Related plasmids used in the study第42-43页
    2.7 Designing primers第43-46页
        2.7.1 Types of Primers第43-44页
        2.7.2 General guidelines第44页
        2.7.3 PCR reaction第44-46页
    2.8 Minipreparation of DNA plasmid第46-47页
    2.9 Electrocompetent cell preparation第47-48页
    2.10 Neurospora electrotransformation第48-50页
    2.11 Genomic DNA preparation第50-51页
    2.12 Neurospora crassa phenotype analysis on the plate medium第51页
    2.13 Race-tube assay第51-52页
    2.14 Neurosspora crassa soluble protein extraction第52-53页
    2.15 SDS-PAGE:Laemmli Method第53-55页
    2.16 Western blot第55-56页
    2.17 NIH Image software for densitometry第56-57页
CHAPTER 3 THE MET-8 GENE EXPRESSION IS REGULATEDBY MULTIPLE FACTORS第57-117页
    3.1 Transcription factors mediate met-8 transcription regulation第57-67页
    3.2 Histone methyltransferase mediate met-8 gene expression第67-73页
    3.3 Histone acethyltransferases regulate met-8 gene expression第73-77页
    3.4 Histone deacetylases participate in met-8 gene regulation第77-83页
    3.5 Chromatin remodeling complexes regulate met-8 gene regulation第83-90页
    3.6 Histone point mutations changed met-8 levels第90-104页
    3.7 Construction of a his-3 targeting vector carrying met-8 promoter第104-105页
    3.8 High-efficiency expression driven by the met-8 promoter第105-107页
    3.9 Steady expression of GFP regulated by the different pmet-8 promoter fragments under difierent conditions第107-108页
    3.10 Sturdy expression of GFP regulated by met-8 promoter under time frame第108-109页
    3.11 The met-8 promoter is capable to drive sustainable expression第109-110页
    3.12 Stable expression of the met-8 promoter第110-112页
    3.13 Independent expression of met-8 under different light intensity第112-114页
    3.14 Independent expression of met-8 promoter under intervallictemperature第114-117页
CHAPTER 4 DISCUSSION第117-120页
REFERENCE第120-127页
Acknowledgements第127-128页
CURRICULUM VITAE第128-129页

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