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The Construction and Application of Biosensor for Detection of Quorum Sensing in Pseudomonas Aeruginosa and Serratia Marcescens

ACKNOWLEDGEMENTS第6-7页
Summary第7-8页
CHAPTER 1:INTRODUCTION AND LITERATURE REVIEW第14-25页
    1.1 SUMMARY OF PSEUDOMONAS AERUGINOSA AND SERRATIA MARCESCENS H30第14-15页
        1.1.1 Summary of Pseudomonas aeruginosa第14页
        1.1.2 Summary of Serratia marcescens H30第14-15页
    1.2 QUORUM SENSING第15-19页
        1.2.1 Summary of quorum sensing第15-16页
        1.2.2 Quorum sensing in Pseudomonas aeruginosa第16-17页
        1.2.3 Quroum sensing in Serratia marcescens第17-18页
        1.2.4 Bacterial cell-cell communication第18-19页
    1.3 AHL SIGNAL第19-23页
        1.3.1 Summary of AHL signal第19-20页
        1.3.2 Extraction of AHL第20页
        1.3.3 Classification of quorum-sensing signals and biological functions第20-21页
        1.3.4 AHLs synthesis第21-22页
        1.3.5 AHL detection第22-23页
            1.3.5.1 Biological detection第22-23页
            1.3.5.2 AHL detection by chemical method第23页
    1.4 OBJECTIVES第23-25页
CHAPTER 2:MATERIALS AND METHODS第25-35页
    2.1 MATERIALS第25-29页
        2.1.1 Bacterial strains第25页
        2.1.2 Growth media第25页
        2.1.3 Plasmid第25-27页
        2.1.4 Oligonucleotide primer第27-28页
        2.1.5 Enzymes第28页
        2.1.6 Miscellaneous and Chemical第28-29页
        2.1.7 Instrument第29页
    2.2 METHODS第29-35页
        2.2.1 Bacterial culture第29-30页
        2.2.2 Primer designation第30页
        2.2.3 Polymerase Chain Reaction (PCR)第30-31页
        2.2.4 PCR product analysis by agarose gel electrophoresis第31页
        2.2.5 Recovery of DNA fragment from agarose gel第31-32页
        2.2.6 Plasmid Extraction第32-33页
        2.2.7 N-Acylhomoserine lactone extraction第33-34页
        2.2.8 Measurement of β-galactosidase activity第34-35页
CHAPTER 3:THE CONSTRUCTION AND APPLICATION OFBIOSENSOR PHL01 FOR THE DETECTION OF LONG-CHAIN AHLS INP.AERUGINOSA ATCC9027第35-47页
    3.1 INTRODUCTION第35页
    3.2 MATERIALS AND METHODS第35-36页
        3.2.1 Bacterial strains and growth conditions第35-36页
        3.2.2 Construction of pHL01 plasmid第36页
    3.3 RESULTS AND DISCUSSION第36-47页
        3.3.1 Construction pHL 01 reporter for detection of long-chain AHL signals第36-42页
        3.3.2 Detecting AHL by using E.coli pHL01 reporter第42-44页
        3.3.3 The influence of some factors on the β-galactosidase activity assay第44-45页
            3.3.3.1 The effect of Culture's pH on the β-galactosidase activity assay第44页
            3.3.3.2 The effect of culture's incubating temperature on the β-galactosidase activityassay第44-45页
            3.3.3.3 The effect of Culture's incubating time on the β-galactosidase activity assay第45页
        3.3.4 Detecting N-acyl homoserine lactones (AHL) from extraction of Pseudomonasaeruginosa第45-47页
CHAPTER 4:THE CONSTRUCTION AND APPLICATION OFBIOSENSOR PHL02 FOR THE DETECTION OF QUORUM SENSING IN S.MARCESCENS H30第47-60页
    4.1 INTRODUCTION第47-48页
    4.2 MATERIALS AND METHODS第48-49页
        4.2.1 Bacterial strains and growth conditions第48页
        4.2.2 Construction of pHL02 plasmid第48-49页
    4.3 RESULTS AND DISCUSSIONS第49-60页
        4.3.1 Construction pHL 02 reporter for detection of long-chain AHL signal第49-53页
        4.3.2 Detection of AHL signal (Sigma)第53-54页
        4.3.3 The influence of some factors on the β-galactosidase activity assay第54-56页
            4.3.3.1 The effect of Culture's pH on the β-galactosidase activity assay第54-55页
            4.3.3.2 The effect of culture's incubating temperature on the β-galactosidase activityassay第55-56页
            4.3.3.3 The effect of Culture's incubating time on the β-galactosidase activity assay第56页
        4.3.4 Detection of AHL from S.marcescens H30's extractions第56-60页
CHAPTER 5:APPLICATION OF E.COLI (PHL02) FOR STUDYING THEEFFECT OF SOME FACTORS ON AHL PRODUCTION FROMPSEUDOMONAS AERUGINOSA AND S. MARCESCENS第60-67页
    5.1 INTRODUCTION第60页
    5.2 MATERIALS AND METHODS第60-61页
        5.2.1 Materials第60-61页
            5.2.1.1 Bacteria第60页
            5.2.1.2 Medium第60-61页
        5.2.2 Methods第61页
            5.2.2.1 Effect of different cultures on AHL production第61页
            5.2.2.2 Influence of temperature on AHL production第61页
            5.2.2.3 Effect of carbon sources on AHL production第61页
            5.2.2.4 Effect of pH of medium on AHL production第61页
    5.3 RESULTS AND DISCUSSION第61-67页
        5.3.1 Effect of different cultures on AHL production第61-62页
        5.3.2 Effect of temperature on AHL production第62-63页
        5.3.3 Effect of pH of medium on AHL production第63-64页
        5.3.4 Effect of carbon sources on AHL production第64-67页
CHAPTER 6:CONCLUSION第67-68页
REFERENCE第68-74页

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