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Analysis of EST Data in the Rice Stem and Using cDNA Microarray to Monitoring Genes Expression Profiles Induced by Gibberellin

DEDICATION第3-4页
DECLARATION第4-5页
CERTIFICATION第5-6页
ACKNOWLEDGMENT第6-7页
LIST OF FIGURES AND TABLES第7-8页
ABBREVIATIONS第8-10页
UNIT ABBREVIATIONS第10-11页
TABLE OF CONTENTS第11-13页
ABSTRACT第13页
Chapter I Introduction and Literature Review第14-31页
    Introduction第14-17页
    Literature Review第17-28页
        1.1 Rice第17-20页
            1.1.1 Aspects about plant growth internodes elongation in stem rice第18-19页
            1.1.2 Some genes that play role in elongation stem in rice plant induced by hormone第19-20页
        1.2 Gibberellins第20-23页
            1.2.1 Functions of Gibberellins第21-22页
            1.2.2 GA-regulated gene expression第22-23页
        1.3 Microarray technology and Expression Sequence Tags第23-28页
            1.3.1 Microarray Technology第23页
            1.3.2 Principle microarray technology第23-24页
            1.3.3 Application第24-25页
            1.3.4 Advantages of microarray第25-26页
            1.3.5 General Aspects about ESTs第26-28页
            1.3.6 Some applications of EST第28页
    Investigation aim and objectives第28-31页
Chapter II. Materials and Methods第31-54页
    2.1 Plant Materials第31页
        2.1.1. GA treatment and samples第31页
    2.2 Extraction of total RNA第31-32页
    2.3 Purification of total RNA第32-33页
    2.4 Isolation of Poly A~+ mRNA from total RNA第33-34页
    2.5 cDNA Library construction第34-40页
        2.5.1 First strand cDNA synthesis第34-36页
        2.5.2 cDNA amplification by LD PCR第36-37页
        2.5.3 Digestion in NotI and SalI第37-38页
        2.5.4 Ligation of ds cDNA to pSPORT II (GIBCO/BRL company) vector第38-39页
        2.5.5 Transformation into E.coli DH5 α第39-40页
    2.6 cDNA library第40-41页
        2.6.1 cDNA library plating第40页
        2.6.2 Clones Inoculation第40-41页
        2.6.3 Plasmid Store (LB/glycerol 1:1)第41页
    2.7 Plasmid extraction by Mini-preparation using Multi-Screen 96-Well Filter plates第41-48页
        2.7.1 Sequencing and analysis第43页
        2.7.2 Preparation of sequencing reaction第43-44页
        2.7.3 Post-reaction clean up第44-45页
        2.7.4 Instruments setup and analysis of data第45页
        2.7.5 Injection and run parameters第45-46页
        2.7.6 Polymerase Chain Reaction (PCR)第46页
        2.7.7 PCR product cleaning第46-47页
        2.7.8 Database building第47页
        2.7.9 ESTs data analysis第47-48页
    2.8 Preparation of cDNA array第48-49页
    2.9 Preparation of labeled cDNA第49页
    2.10 Chip hybridization第49-51页
        2.10.1 Prehybridization第49-50页
        2.10.2 Hybridization第50页
        2.10.3 Membranes washing第50-51页
    2.11 Scanning the membrane第51页
    2.12 Quantitative real-time PCR第51-54页
Chapter III Results第54-68页
    3.1 Characterization of the cDNA library第54-56页
    3.2 Identification analysis of TUTs sequences and similarities found第56页
    3.3 Functional annotation第56-59页
    3.4 Highly expressed genes第59-61页
    3.5 Homologous comparison to other organism第61-62页
    3.6 Analysis of cDNA array data and identification of differentially expressed genes第62-66页
    3.7 Quantitative real-time PCR analysis to validate array results第66-68页
Chapter IV. Discussion and conclusions第68-86页
    4.1 High genes expression profiles of the rice stem relate to ESTs functional annotation第68-71页
    4.2 Genes with high expression library induced by gibberellin in 36h第71-74页
    4.3 Differentially expressed genes related with metabolic process induced by gibberellin in 12 and 36 h第74-85页
    4.4 Conclusion第85-86页
Recommendation第86-87页
Reference List第87-99页

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