共表达PCV2 Cap基因及TGEV S基因的真核重组质粒构建及实验免疫研究

发布时间：2021-05-18 12:18

　　猪圆环病毒属于环状病毒科，圆环病毒属，其成员包括PCV1和PCV2。PCV1对猪没有致病性，而PCV2可以引起断奶仔猪多系统衰竭综合征（PMWS）。在国产猪中PCV2的感染非常普遍，而且在五大洲各个国家都诊断出有PMWS发生，患病率在4-30%不等。猪传染性胃肠炎病毒（TGEV）是单链RNA病毒，属于尼多病毒目、冠状病毒科，能够引起猪的传染性胃肠炎。各个品种和各个年龄段的猪都对TGEV易感，并且在感染后3周内死亡率可达到100%。因此，PCV2和TGEV在世界范围内的养猪业中都是毁灭性疾病，预防接种是避免PCV2和TGEV感染的有效方法。虽然现有的抗PCV2和TGEV的疫苗可以有效地减少临床症状的出现，并且能够提高农场生产量，但这些疫苗无法完全防止PCV2和猪TGEV的感染和传播。此外，一些实验性的灭活疫苗、DNA疫苗、含有PCV2Cap或TGEV S基因的重组活病毒载体疫苗也用于抗PCV2和TGEV的感染。尽管如此，这是首次在同一种疫苗载体中共表达PCV2Cap和TGEV S基因，并用其免疫猪以抵抗PCV2和TGEV的感染。本研究的目的是在真核表达载体中共表达PCV2Cap和TGEV... 

【文章来源】：吉林大学吉林省 211工程院校 985工程院校 教育部直属院校

【文章页数】：156 页

【学位级别】：博士

【文章目录】：
中文摘要
Abstract
List of Abbreviations
Foreword
Chapter 1 Literature Review
    1.1 Introduction of PCV2 and (PMWS)
        1.1.1 historical of porcine circovirus type 2
        1.1.2 General epidemiology
        1.1.3 Transmission
        1.1.4 Disease and Syndrome
        1.1.5 Diagnosis
        1.1.6 Prevention and control of PCV2
    1.2 Introduction of PCV
        1.2.1 Evolution of PCV
        1.2.2 Viral genome structure
        1.2.3 Structural proteins
        1.2.4 Virus replication
        1.2.5 Host Innate Immune Response against Viral Infection
        1.2.6 Cells mediated
        1.2.7 Neutralization Antibody (NA)
        1.2.8 Vaccine development against PCV2 infection
    1.3 Transmissible gastroenteritis
        1.3.1 Introduction of TGEV
        1.3.2 Etiology
        1.3.3 Epidemology
        1.3.4 Disease and syndrome
        1.3.5 Diagnostic
        1.3.6 TGEV viral structure
        1.3.7 Genome organization and antigenic relationships
        1.3.8 Immunity
        1.3.9 Progress in genetically engineered vaccines against TGE-Virus
    1.4 Objective of this study
Chapter 2 construction of recombinant plasmids and the proteins expression in eukaryotic cells
    2.1 Introduction
    2.2 plasmids construction design
    2.3 Materials and methods
        2.3.1 Materials
        2.3.2 Methods
    2.4 Results
        2.4.1 pMD-18T-Cap and pMD-18T-S plasmids construction
        2.4.2 Sequencing and sequence analysis
        2.4.3 pEGFP-ORF2 plasmid construction
        2.4.4 pEGFP-Cap, pEGFP-S and pEGFP-Cap-S plasmids transfection and expressed fused protein identification
        2.4.5 Indirect ELISA
    2.5 Discussion
        2.5.1 Gene fragments design and plasmids construction
        2.5.2 Transfection and fused proteins identification
        2.5.3 IFN-γ detection in transfected PK-15 cells
    2.6 Summary
Chapter 3 Characterization of immune responses to vaccination and infection in pigs75
    3.1 Introduction
    3.2 Materials and Methods
        3.2.1 Materials
        3.2.2 Methods
    3.3 Results
        3.3.1 Humoral immune responses in pigs immunized with DNA vaccines
        3.3.2 Pigs survival rate after TGEV challenge (%)
        3.3.3 Cell immunity
    3.4 Discussion
        3.4.1 Humoral immune response
        3.4.2 Cells immune responses
    3.5 Summary:
Chapter 4 Histopathological analysis in immunized and PCV2 or TGEV challenged pigs
    4.1 Introduction
    4.2 Materials and Methods
        4.2.1 Material
        4.2.2 Methods
    4.3 Results
        4.3.1 Histopathology examination
        4.3.2 Immunohistochemistry (IHC)
    4.4 Discussion
        4.4.1 HE
        4.4.2 IHC
    4.5 Summary
Conclusion
References
导师简介
Curriculum vitae
攻读博士期间发表的学术论文及其它成果
致谢


【参考文献】：
期刊论文
[1]Comparison of Immune Responses against FMD by a DNA Vaccine Encoding the FMDV/O/IRN/2007 VP1 Gene and the Conventional Inactivated Vaccine in an Animal Model[J]. Farahnaz Motamedi Sedeh,Hoorieh Soleimanjahi,AmirReza Jalilian,Homayoon Mahravani.  Virologica Sinica. 2012(05)



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