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家蚕来源的一株单链RNA病毒的分离和特性研究

DEDICATION第4-5页
ACKNOWLEDGEMENT第5-7页
DECLARATION第7-8页
CERTIFICATION第8-14页
Abstract第14页
General Introduction第18-36页
Literature review第36-55页
Chapter 1 Virus source, multiplication, Isolation, purification, and Electron Microscopic study of the virus第55-65页
    1 Introduction第55-65页
        1.1 Life cycle of silkworms第55-57页
        1.2 Materials第57-58页
        1.3 Methods第58-60页
            1.3.1 Procurement of Experimental material第58页
            1.3.2 Virus source第58-59页
            1.3.3 Method of virus infection第59页
            1.3.4 Extraction of crude virus from diseased silkworms第59-60页
        1.4 Virus purification第60-61页
        1.5 Transmission Electron Microscopy第61页
        1.6 Results and Discussion第61-65页
Chapter 2 Preparation of Polyclonal antibodies, Histopathological studies of the infected silkworms by Immunofluoresence study and Pyronine staining of the virus infected tissues第65-79页
    2 Production of Polyclonal Antibodies第66-79页
        2.1 Introduction第66-67页
        2.2 Materials第67页
        2.3 Methods第67-68页
        2.4 Immunofluoresence study第68-72页
            2.4.1 Introduction第68-69页
            2.4.2 Materials第69页
            2.4.3 Methods第69-72页
                2.4.3.1 Artificial induction of the disease第69页
                2.4.3.2 Sterilization of the equipments第69-70页
                2.4.3.3 Processing of infected tissues for Immunofluoresence studies第70页
                2.4.3.4 Sectioning samples in wax blocks第70-71页
                2.4.3.5 Immunofloresence staining第71-72页
        2.5 Results and Discussion第72-77页
        2.6 Pyronine Methyl Green staining第77-78页
            2.6.1 Introduction第77页
            2.6.2 Materials第77页
            2.6.3 Methods第77-78页
                2.6.3.1 Reproduction of Methyl Green Pyronine stain第77-78页
                2.6.3.2 Staining method第78页
        2.7 Results and Discussion第78-79页
Chapter 3 Isolation and determination of Nucleic acid of the virus by Formaldehyde test and Gel Immunodiffusion test第79-87页
    3 Extraction of RNA from virus第80-87页
        3.1 Introduction第80-81页
        3.2 Materials第81页
        3.3 Methods第81-82页
            3.3.1 Sterilization of Glassware and Equipment第81页
            3.3.2 Extraction of RNA第81-82页
        3.4 Analysis of virus Genome for DNA and RNA第82页
            3.4.1 Introduction第82页
                3.4.1.1 Diphenylamine test第82页
                3.4.1.2 Orcinol test第82页
        3.5 Materials第82页
        3.6 Methods第82-83页
        3.7 Results and Discussion第83页
        3.8 Gel Immtmodiffusion Test第83-84页
        3.9 Materials第84页
        3.10 Methods第84页
        3.11 Results and Discussion第84-85页
        3.12 Formaldehyde test第85页
            3.12.1 Introduction第85页
        3.13 Materials第85页
        3.14 Methods第85页
        3.15 Results and Discussion第85-87页
Chapter 4 SDS-PAGE profile of the virus coat protein, Detection of virus by PCR, Partial sequencing of viral Genome第87-115页
    4 SDS PAGE Electrophoresis第88-115页
        4.1 Introduction第88-89页
            4.1.1 Separation of proteins第88-89页
        4.2 SDS PAGE of proteins第89-94页
            4.2.1 Separation of proteins under denaturing conditions第89页
            4.2.2 Determination of Molecular weights of proteins第89页
            4.2.3 Continuous and discontinuous buffer systems第89-90页
            4.2.4 Materials第90-91页
            4.2.5 Methods第91-94页
                4.2.5.1 Reagents used in Gels第91-92页
                4.2.5.2 SDS PAGE of virus proteins第92页
                4.2.5.3 Preparation of protein sample第92-93页
                4.2.5.4 Electrophoresis第93-94页
        4.3 Results and Discussion第94-97页
        4.4 Detection of virus by PCR第97-99页
            4.4.1 Introduction第97-99页
        4.5 RT-PCR第99页
        4.6 Materials第99页
        4.7 Methods第99-100页
            4.7.1 Extraction of RNA from the virus第99-100页
        4.8 PCR第100-101页
            4.8.1 Positive control RNA one step RT-PCR第100-101页
        4.9 Nested RT-PCR第101-102页
        4.10 Results and Discussion第102-104页
        4.11 Partial Genome sequencing of the virus第104-105页
            4.11.1 Introduction第104-105页
        4.12 Materials第105-106页
        4.13 Methods第106-107页
            4.13.1 One step RT-PCR第106-107页
        4.14 Results and Discussion第107-115页
5 References第115-128页

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