共表達(dá)PCV2 Cap基因及TGEV S基因的真核重組質(zhì)粒構(gòu)建及實(shí)驗(yàn)免疫研究
發(fā)布時(shí)間:2021-05-18 12:18
豬圓環(huán)病毒屬于環(huán)狀病毒科,圓環(huán)病毒屬,其成員包括PCV1和PCV2。PCV1對(duì)豬沒(méi)有致病性,而PCV2可以引起斷奶仔豬多系統(tǒng)衰竭綜合征(PMWS)。在國(guó)產(chǎn)豬中PCV2的感染非常普遍,而且在五大洲各個(gè)國(guó)家都診斷出有PMWS發(fā)生,患病率在4-30%不等。豬傳染性胃腸炎病毒(TGEV)是單鏈RNA病毒,屬于尼多病毒目、冠狀病毒科,能夠引起豬的傳染性胃腸炎。各個(gè)品種和各個(gè)年齡段的豬都對(duì)TGEV易感,并且在感染后3周內(nèi)死亡率可達(dá)到100%。因此,PCV2和TGEV在世界范圍內(nèi)的養(yǎng)豬業(yè)中都是毀滅性疾病,預(yù)防接種是避免PCV2和TGEV感染的有效方法。雖然現(xiàn)有的抗PCV2和TGEV的疫苗可以有效地減少臨床癥狀的出現(xiàn),并且能夠提高農(nóng)場(chǎng)生產(chǎn)量,但這些疫苗無(wú)法完全防止PCV2和豬TGEV的感染和傳播。此外,一些實(shí)驗(yàn)性的滅活疫苗、DNA疫苗、含有PCV2Cap或TGEV S基因的重組活病毒載體疫苗也用于抗PCV2和TGEV的感染。盡管如此,這是首次在同一種疫苗載體中共表達(dá)PCV2Cap和TGEV S基因,并用其免疫豬以抵抗PCV2和TGEV的感染。本研究的目的是在真核表達(dá)載體中共表達(dá)PCV2Cap和TGEV...
【文章來(lái)源】:吉林大學(xué)吉林省 211工程院校 985工程院校 教育部直屬院校
【文章頁(yè)數(shù)】:156 頁(yè)
【學(xué)位級(jí)別】:博士
【文章目錄】:
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
導(dǎo)師簡(jiǎn)介
Curriculum vitae
攻讀博士期間發(fā)表的學(xué)術(shù)論文及其它成果
致謝
【參考文獻(xiàn)】:
期刊論文
[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)
本文編號(hào):3193776
【文章來(lái)源】:吉林大學(xué)吉林省 211工程院校 985工程院校 教育部直屬院校
【文章頁(yè)數(shù)】:156 頁(yè)
【學(xué)位級(jí)別】:博士
【文章目錄】:
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
導(dǎo)師簡(jiǎn)介
Curriculum vitae
攻讀博士期間發(fā)表的學(xué)術(shù)論文及其它成果
致謝
【參考文獻(xiàn)】:
期刊論文
[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)
本文編號(hào):3193776
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