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Study of a New C-type Lectin with a Long Carbohydrate Recognition Domain(CRD)from Artemia Parthenogenetica

ACKNOWLEDGEMENTS第4-7页
ABSTRACT第7页
I INTRODUCTION第8-11页
II LITERATURE第11-20页
    II 1.ANIMAL LECTINS第12-20页
        II 1. 1. DETECTION第12-13页
        II.1.2 ISOLATION AND PURIFICATION第13页
        II.1.3. CLASSIFICATION第13-16页
            II.1.3.1. Galectins第13-14页
            II.1.3.2. C-type lectins第14页
            II.1.3.3. P-type lectins第14页
            II.1.3.4. Pentraxins第14-15页
            II.1.3.5. Siglecs第15页
            II.1.3.6. Calnexin,calreticulin and calmegin第15页
            II.1.3.7. Ficolins and intelectins第15-16页
            II.1.3.8. Other animal lectins第16页
        II.1.4. FUNCTIONS第16-18页
        II.1.5. APPLICATIONS第18-20页
III MATERIALS AND METHODS第20-40页
    III.1. ANIMALS第21页
    III.2. CLONING OF THE FULL LENGTH cDNA ENCODING ApLC第21-24页
        III.2.1. PURIFICATION OF RNA第21-22页
        III.2.2. QUANTIFICATION OF RNA/DNA第22页
        III.2.3. RT-PCR FOR PREPARATION OF FIRST STRAND cDNA第22-23页
        III.2.4. AMPLIFICATION OF ApLC FULL FROM FIRST STRAND cDNA第23-24页
            III.2.4.1. PRIMER DESIGN AND PREPARATION第23-24页
            III.2.4.2. PCR COMPONENTS第24页
            III.2.4.3. PCR CONDITIONS第24页
    III.3. PURIFICATION OF THE AMPLIFIED cDNA第24-25页
    III.4. LIGATION OF ApLC第25页
    III.5. TRANSFORMATION第25-26页
    III.6. INSERT CHECK AND CLONING OF THE TRANSFORMED E.COLI第26页
    III.7. SEQUENCE ANALYSIS AND PHYLOGENETIC TREE CONSTRUCTION第26-27页
    III.8. FAR SOUTHERN BLOTTING第27-31页
        III.8.1. AMPLIFICATION AND LABELING OF THE PROBE第27页
        III.8.2. PREPARATION OF SOLUTIONS FOR FAR-SOUTHEN BLOTTING第27-29页
        III.8.3. PERFORMING THE FAR-SOUTHERN BLOTTING第29-31页
            III.8.3.1. Washings of loaded gel第30页
            III.8.3.2. Transferring of DNA onto membrane and probe hybridization第30页
            III.8.3.3. Washing of membrane and signal detection第30-31页
    III.9. RECOMBINANT EXPRESSION OF ApLC第31-34页
        III.9.1. Construction of the expression plasmid第31页
        III.9.2. Expression of ApLC第31-32页
        III.9.3. Purification of ApLC第32-34页
            III.9.3.1. Preparation of buffers第32-33页
            III.9.3.2. Purification第33-34页
    III.10. SDS-Polyacrylamide Electrophoresis(SDS-PAGE)第34-37页
        III.10.1. Reagents preparation第34-35页
        III.10.2. Performing the SDS-Polyacrylamide Gel Electrophoresis第35-37页
    III.11. HEMAGGLUTINATION ASSAY第37-38页
    III.12. STUDY OF FACTORS MODULATING ApLC ACTIVITY第38页
    III.13. CARBOHYDRATES INDIRECT BINDING ASSAY OF ApLC第38-39页
    III.14. BACTERIAAGGLUTINATION TEST第39页
    III.15. ANTIMICROBIAL ACTIVITY ASSAY第39-40页
IV. RESULTS第40-67页
    IV 1.cDNA CLONING AND SEQUENCE ANALYSIS OF ApLC第41-43页
    IV 2.HOMOLOGOUS AND PHYLOGENETIC ANALYSIS OF ApLC第43-45页
    IV 3.BODYPARTS EXPRESSION OF ApLC第45-47页
    IV.4. EXPRESSION OF ApLC AFTER BACTERIA CHALLENGE第47-53页
    IV.5. EXPRESSION AND PURIFICATION OF RECOMBINANT ApLC第53-54页
    IV.6. HEMAGGLUTINATION ASSAY第54-55页
    IV 7.SUGAR SPECIFICITY ASSAY第55页
    IV.8. DEPENDANCE OF ApLC ACTIVITY ON PH DIVALENT CATIONS AND TEMPERATURE第55-57页
    IV 9.BACTERIA-AGGLUTINATION ACTIVITY OF ApLC第57-62页
    IV 10.ANTIMICROBIAL ACTIVITY OF ApLC第62-67页
VI. CONCLUSION第67-69页
VII. REFERENCES第69-79页

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