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Molecular Epidemiology of Piroplasms in Pakistan, and Molecular Characterization, and Functional Analysis of HSP90Proteins of Babesia Orientalis

Contents第4-7页
Abbreviations第7-10页
Abstract第10-11页
Chapter 1-Review of Literature第14-43页
    1.1 Babesia and Theileria第14-34页
        1.1.1 General Introduction第14页
        1.1.2 Life cycle第14-17页
        1.1.3 Pathophysiology and clinical manifestation第17-20页
        1.1.4 Epidemiology第20-28页
            1.1.4.1 Geographical distribution第20-26页
            1.1.4.2 Transmission dynamics第26-28页
        1.1.5 Advances in diagnosis第28-33页
            1.1.5.1 Various types of PCR第29-30页
            1.1.5.2 Loop mediated isothermal amplification assay第30页
            1.1.5.3 Reverse line blot hybridization assay第30-31页
            1.1.5.4 Role of molecular biology in the detection of new species第31-33页
        1.1.6 Conclusions and hypothesis第33-34页
    1.2 Heat Shock Protein 90(HSP90)第34-43页
        1.2.1 General Introduction第34页
        1.2.2 Structure of HSP90第34-40页
            1.2.2.1 N-terminal domain第36-38页
            1.2.2.2 Middle domain第38-39页
            1.2.2.3 C-terminal domain第39-40页
        1.2.3 Function of HSP90第40-41页
        1.2.4 HSP90 in apicomplexan parasites第41-42页
        1.2.5 Conclusions and hypothesis第42-43页
Chapter 2-Molecular Epidemiology of Theileria annulata in Pakistan第43-66页
    2.1 Introduction第43-44页
    2.2 Materials and methods第44-54页
        2.2.1 Study area and sample collection第44-46页
        2.2.2 Epidemiological survey第46页
        2.2.3 DNA extraction第46页
        2.2.4 Reverse Line Blot Hybridization assay第46-51页
            2.2.4.1 Polymerase chain reaction第46-48页
            2.2.4.2 Preparation of membrane第48-50页
            2.2.4.3 Reverse line blot hybridization第50-51页
        2.2.5 Cloning and sequencing of 18S rRNA gene and ITS regions第51-53页
        2.2.6 Phylogenetic and sequence analysis第53-54页
        2.2.7 Statistical analysis第54页
    2.3 Results第54-62页
        2.3.1 Prevalence and RLB results第54页
        2.3.2 Nucleotide sequence heterogeneity analysis第54-58页
        2.3.3 Phylogenetic analysis第58-59页
        2.3.4 Associated risk factors第59-62页
    2.4 Discussion第62-66页
Chapter 3-Two Putative HSP90 Proteins in Babesia orientalis第66-97页
    3.1 Introduction第66-67页
    3.2 Materials and methods第67-74页
        3.2.1 In silico identification of two HSP90-like proteins of B. orientalis第67页
        3.2.2 Parasites and animals第67页
        3.2.3 DNA and RNA extraction and preparation of cDNA第67-68页
        3.2.4 Amplification,cloning and sequencing of BoHSP90-A and BoHSP90-B genes第68-69页
        3.2.5 Sequence structure,function and motifs prediction第69-70页
        3.2.6 Recombinant expression and purification of BoHSP90-A and BoHSP90-B第70-72页
        3.2.7 Production of anti-rBoHSP90-A and anti-rBoHSP90-B immune serum第72-73页
        3.2.8 SDS-PAGE and western blotting第73-74页
    3.3 Results第74-93页
        3.3.1 Molecular characterization and sequence analysis第74-76页
        3.3.2 Predicted structure and functional analysis第76-82页
        3.3.3 HSP90 inhibitors第82-88页
        3.3.4 Antigenicity analysis第88-91页
        3.3.5 Identification of BoHSP90-A and BoHSP90-B第91-93页
    3.4 Discussion第93-97页
Chapter 4-Conclusions and Future Prospects第97-99页
References第99-123页
List of Tables第123-124页
List of Figures第124-126页
APPENDICES第126-127页
ACKNOWLEDGEMENT第127-129页
PUBLICATIONS第129-130页

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