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Identification and Characterization of Cis-Encoded Antisense RNAs Associated with the Replication Process of Salmonella Enterica Serovar Typhi

ABSTRACT第7-9页
摘要第10-15页
ABBREVIATIONS第15-17页
CHAPTER 1 Introduction第17-35页
    1.1 Background第17页
    1.2 Salmonella第17-19页
    1.3 Bacterial non-coding RNAs第19页
    1.4 Bacterial antisense RNAs第19-25页
        1.4.1 Characteristics of cis-encoded antisense RNAs第20页
        1.4.2 Mechanisms of action of cis-encoded antisense RNAs in bacteria第20-22页
        1.4.3 Methods to find asRNAs第22-25页
    1.5 DNA Replication in Bacteria第25-27页
        1.5.1 Replication origin architecture第26页
        1.5.2 Overview of bacterial replication initiation第26-27页
        1.5.3 Elongation of DNA第27页
        1.5.4 Termination of DNA replication第27页
    1.6 Objectives第27-28页
    1.7 Relevance of the study第28页
    1.8 Experimental design第28-30页
    1.9 References第30-35页
CHAPTER 2 An antisense RNA AsdA increases the mRNA stability of the gene encoding the replicationinitiation protein of S. Typhi第35-75页
    2.1 Introduction第35-36页
    2.2 Materials and Methods第36-46页
        2.2.1 Bacterial strains,plasmids,and growth conditions第36-37页
        2.2.2 Construction of strains overexpressing the asRNAs第37-40页
        2.2.3 Construction of strains overexpressing the asRNAs in RNase Ⅲ and RNase E mutants第40页
        2.2.4 Construction of fur mutant第40页
        2.2.5 5'-RACE第40-41页
        2.2.6 3'-RACE第41-42页
        2.2.7 Growth kinetic analysis第42-43页
        2.2.8 Expression analysis of asRNA under stress conditions第43页
        2.2.9 Overexpression analysis第43页
        2.2.10 RNA extraction第43-44页
        2.2.11 Quantitative RT-PCR第44页
        2.2.12 Northern blot analysis第44页
        2.2.13 Growth curves第44-46页
    2.3 Results第46-66页
        2.3.1 Identification and mapping of the 5' and 3' ends of AsdA第46-47页
        2.3.2 Analysis of AsdA expression under different growth conditions第47-50页
        2.3.3 Strain constructions第50-61页
        2.3.4 Effect of overexpression of asdA on dnaA mRNA level第61-63页
        2.3.5 Effect of overexpression of asdA on the growth of S. Typhi第63-66页
    2.4 Discussion第66-70页
    2.5 References第70-75页
CHAPTER 3 Identification and characterization of a cis antisense RNA of parC gene encoding DNAtopoisomerase Ⅳ of S. Typhi第75-97页
    3.1 Introduction第75-77页
    3.2 Materials and Methods第77-81页
        3.2.1 Bacterial strains and culture conditions第77页
        3.2.2 Strain and plasmid construction第77-78页
        3.2.3 5'-and 3'-RACE第78页
        3.2.4 RNA extraction第78-79页
        3.2.5 Northern blot analysis第79页
        3.2.6 Quantitative RT-PCR第79-80页
        3.2.7 Growth curves第80页
        3.2.8 Motility assay第80-81页
    3.3 Results第81-90页
        3.3.1 Identification of antisense RNA complementary to parC mRNA第81-82页
        3.3.2 Expression of AspC under different growth conditions第82-84页
        3.3.3 Strain constructions第84-85页
        3.3.4 Effect of overexpression of AspC on parC mRNA level第85-88页
        3.3.5 Effect of overexpression of AspC on the growth of S. Typhi第88-90页
    3.4 Discussion第90-94页
    3.5 References第94-97页
GENERAL CONCLUSIONS第97-99页
ACKNOWLEDGEMENTS第99-101页
LIST OF PUBLICATIONS第101-103页
APPENDICES第103-105页

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