The concept and history of 1. 1 vaccine

Application and effect analysis of 1.2 vaccine 3

1.3 development of new technologies for vaccine research 6

1.3. 1 traditional vaccine 7

1.3.2 genetically engineered vaccine 7

1.4 development prospect of vaccine in eliminating and controlling animal infectious diseases 8 pages

Reference 8

Chapter II Basic Theory of Vaccine Immunology Page 9

2. 1 fundamentals of vaccine-related immunology 9

2. 1. 1 immune system 9

2. 1.2 immune organs 9

Immune cells 1 1

2. 1.4 antigen 13

2. 1.5 antibody 15

2. 1.6 Basic structure and function of immunoglobulin 16

2.2 Basic process of immune response 17

2.2. 1 nonspecific immune response 17

2.2.2 Specific immune response 19

2.3 Basic elements of effective immune response of vaccine 22

2.3. 1 antigen factor 22

Physical factors 22

2.3.3 Impact of immunization methods 23

2.4 Active immune response of vaccine immunization 23

2.4. 1 antigen presentation 23

2.4.2 antigen competition

2.4.3 Dynamic changes of humoral immunity and cellular immune response 25

2.4.4 Regulation of immune response 25

2.4.5 Immune memory and immune promotion 26

2.4.6 Systemic immunity, local immunity and colostrum immunity 27

Passive immunization 28

Reference 28

Chapter III Vaccine Adjuvants 30

3. 1 Mechanism of action of vaccine adjuvant 30

3. 1. 1 regulating immunity 30

3. 1.2 presenting antigen 3 1

3. 1.3 Cytotoxic T cell reaction 3 1

3. 1.4 storage effect 32

3.2 Plant Additives 32

3.2. 1 saponin 32

3.2.2 Immune Stimulation Compound Adjuvant 32

Propolis adjuvant 33

Lentinan 34

3.2.5 Coriolus versicolor polysaccharide 34

3.2.6 Astragalus polysaccharide 34

3.3 Bacterial Adjuvants 35

3.3. 1 lipopolysaccharide 35

3.3.2 Cell wall acyl dipeptide and its derivatives 35

Cholera toxin 35

3.3.4 Escherichia coli heat-labile enterotoxin 36

Pertussis toxin 36

3.3.6 Corynebacterium parvum 36

BCG 37

3.3.8 Monophosphoryl lipid A37

3.4 Mineral oil additives 37

3.4. 1 oil emulsion 37

MF59 adjuvant 37

3.5 mineral salt additives 38

3.5. 1 aluminum adjuvant (aluminum glue) 38

3.5.2 Calcium phosphate adjuvant 39

3.5.3 Iron hydroxide gel adjuvant 40

Selenium 40

3.6 cytokine adjuvant 40

3.6. 1 interleukin? 14 1

3.6.2 Interleukin? 24 1

3.6.3 Interleukin? 124 1

3.6.4 Granulocytes? Macrophage colony stimulating factor 4 1

Interferon 42

3.6.6 Other cytokines

3.7 nucleic acid adjuvant 43

3.7. 1 immunostimulatory sequence (CpG motif) 43

3.7.2 Nucleic acid vector expressing immune-related cytokines 46

3.7.3 double-stranded RNA46

3.8 Conveying system 47

3.9 Selection and research direction of new adjuvants 47

Reference 49

Chapter IV Technical Basis of Vaccine Design 5 1

4. 1 Technical characteristics of classical vaccines and new vaccines; 5 1

4. 1. 1 Technical Points of Classic Vaccine 5 1

4. 1.2 characteristics of the new vaccine 52

4.2 Current trends in vaccine research 52

4.2. 1 Molecular design of new vaccine 52

4.2.2 Mass production of new vaccines 53

4.2.3 Development of matching differential diagnosis technology 53

4.2.4 Use of new adjuvants 53

4.2.5 Construction of Veterinary Biological Products Engineering Research Center

4.3 Research and development principles of routine live vaccine 53

4.3. 1 natural attenuated strain of pathogen 53

4.3.2 Heterologous Immunity 53

4.3.3 Passage weakening of heterologous animals or cells 54

4.3.4 Changing the external environment for generations 54

4.4 Genetic engineering vaccine and its research and development principles 54

4.4. 1 subunit vaccine and its research and development principles 54

4.4.2 Gene deletion vaccine and its research and development principles 55

4.4.3 Live vector vaccine and its research and development principles 55

4.4.4 Nucleic acid vaccine and its research and development principles 56

4.4.5 Synthetic peptide vaccine and its research and development principles 58

4.4.6 T cell vaccine 60

4.5 Key factors for optimizing gene expression in genetic engineering vaccine design 6 1

4.5. 1 codon optimization 1

4.5.2 Translation termination efficiency 62

4.5.3 Expression of heterologous protein in eukaryotic cells 62

4.6 Computer Aided Vaccine Design Technology 63

4.6. 1 Principles and methods of computer-aided vaccine design technology 63

4.6.2 Application of Computer Aided Vaccine Design Technology and Common Tools 64

4.6.3 Industrial Prospect of Computer Aided Vaccine Design 65

4.7 Screening of highly effective vaccine by immunoprotein histology 65

4.7. 1 Immunoprotein omics

4.7.2 Technical system of immune protein omics 66

4.7.3 Application of Immunoprotein Genomics in Screening Vaccine Candidate Targets 67

Outlook 67

Reference 68

Chapter V Immunological Evaluation of Vaccine Effect 69

5. 1 principles and steps of vaccine development 69

5.2 General principles of immunological evaluation of vaccine efficacy 69

5.2. 1 method 69

Safety 70

Immune effect 70

Protective effect 70

5.2.5 Epidemiological assessment

5.3 Immunological evaluation method of vaccine effect 70

5.3. 1 Laboratory evaluation of vaccine immune effect 70

5.3.2 Clinical evaluation of vaccine immune effect

5.4 Epidemiological evaluation of vaccine effect 74

5.4. 1 Infectious disease epidemic situation and vaccine action strategy 74

5.4.2 Prevalence and epidemiological characteristics of infectious diseases 74

5.4.3 The role of vaccines in controlling the epidemic of infectious diseases 76

5.4.4 Vaccine immunization strategy 76

5.4.5 Epidemiological indicators of vaccine efficacy 77

5.4.6 Epidemiological design of vaccine effect investigation 78

Reference 79

Chapter VI Overview of Genetic Engineering Vaccine 8 1

6. 1 the concept of genetic engineering vaccine

6.2 Genetic engineering subunit vaccine 82

6.2. 1 bacterial disease subunit vaccine 82

6.2.2 Virus disease subunit vaccine 83

6.2.3 Hormone subunit vaccine 83

6.3 Gene mutation vaccine and gene deletion vaccine 83

6.4 Genetic engineering live vector vaccine 84

6.4. 1 Replicated live vector vaccine 84

Non-replicating vector vaccine 85

6.5 nucleic acid vaccine 85

6.6 Edible vaccines for transgenic plants 85

6.7 Synthetic peptide vaccine 86

6.8 Anti-idiotypic vaccine 87

6.9 Development Status and Prospect of Genetic Engineering Vaccine Industry for Livestock and Poultry 87

6.9. 1 International industrialization of genetic engineering vaccines for livestock and poultry 88

6.9.2 Domestic Industrialization Status of Genetic Engineering Vaccines for Livestock and Poultry 88

6. 9. 3 2 1 Opportunities, Challenges and Strategies Faced by Century 89

Reference 90

Chapter VII Genetic Engineering Subunit Vaccine 92

7. 1 Types of genetically engineered subunit vaccines 92

7.2 Genetic engineering subunit vaccine antigen gene selection 93

7.3 antigen expression system of genetically engineered subunit vaccine 93

7.3. 1 prokaryotic expression system 93

7.3.2 Yeast Expression System 96

7.3.3 insect cell expression system 10 1

7.3.4 Mammalian Cell Expression System 104

7.4 Example of Exogenous Protein Expression 1 10

7.4. 1 Prokaryotic expression of foreign protein 1 10

7.4.2 Pichia pastoris expressed foreign protein 1 1 1.

7.4.3 Insect cells (Bac? Where to? Bac system) expressed foreign protein 1 14.

7.4.4 Transfection of Eukaryotic Cells 1 15 by Liposome Method

7.5 Research and application of subunit vaccine 1 15

7.5. 1 Application of prokaryotic expression system in subunit vaccine research 1 15

7.5.2 Application of eukaryotic expression system in subunit vaccine research 1 16

7.6 Outlook 1 17

Reference 1 17

Chapter VIII Transgenic Plant Vaccine 1 19

8. 1 Research progress in the production of edible vaccines by using plant bioreactors 120

8. 1. 1 Expression System of Edible Vaccine 5438+020

8. 1.2 Feasibility of edible vaccine 12 1

8.2 Vector System of Transgenic Plant Vaccine 123

8.2. 1 Plant virus vector with foreign gene transferred 123

8.2.2 Plasmid vector 125 for foreign gene transfer

8.2.3 Carrier Card Box 130

8.2.4 Selection marker genes and reporter genes commonly used in vector construction 13 1

8.2.5 Transformation method of foreign gene 135

8.2.6 Status and expression of transgene and gene silencing 145

8.3 Plant Receptor System of Transgenic Plant Vaccine 148

8.3. 1 Types and characteristics of plant gene transformation receptor system 148

8.3.2 Procedure for Establishing Plant Gene Transformation Receptor System 149

8.3.3 Common Problems in the Establishment of Plant Gene Transformation Receptor System 152

8.4 transgenic plants 153 operation case

8.4. 1 Agrobacterium tumefaciens-mediated gene transformation method 153

8.4.2 Agrobacterium rhizogenes mediated plant transformation 16 1

8.4.3 Gene gun bombardment method (instantaneous expression) 163

8.5 Problems and Countermeasures of Transgenic Plant Vaccine 163

8.5. 1 recipient plant is not ideal 164

8.5.2 The expression level of recombinant antigen protein is low 164.

8.5.3 The immunogenicity of transgenic plant vaccine is weak 164.

8.5.4 Security Issues 165

8.5.5 When a large number of foreign genes are expressed in plants, their growth appears weak. 165.

8.5.6 Digestion and degradation of antigen during oral administration 165

8.5.7 Purification of Recombinant Protein 165

8.5.8 Immunotolerance 165

8.6 Potential customers 166

Reference 166

Chapter 9 Virus Gene Deletion Vaccine 169

9. 1 investigation of gene deletion vaccine 169

9.2 Preparation principle and technology of gene deletion vaccine for livestock and poultry 17 1

9.2. 1 preparation principle and method of herpes virus gene deletion vaccine 17 1

9.2.2 Preparation Principle and Method of Retrovirus Gene Deletion Vaccine 175

9.2.3 preparation principle and method of RNA virus gene deletion vaccine 175

9.3 Research and Application Status of Gene Deletion Vaccine for Livestock and Poultry 180

9.3. Study on1Pseudorabies Gene Deletion Vaccine 180

9.3.2 Study on Gene Deletion Vaccine for Bovine Infectious Rhinotracheitis 180

9.3.3 Study on equine herpesvirus gene deletion vaccine 18 1

9.3.4 Study on Retrovirus Gene Deletion Vaccine 18 1

9.3.5 Study on RNA virus gene deletion vaccine 18 1

9.3.6 Application Status of Gene Deletion Vaccine in Livestock and Poultry 182

9.4 Prospect of Gene Deletion Vaccine for Livestock and Poultry 182

Reference 183

Chapter 10 virus live vector vaccine 186

10. 1 technical characteristics of recombinant virus live vector vaccine 5438+086

10.2 design principles of recombinant live vector vaccine 187

Select 10.2. 1 antigen 187.

10.2.2 selection of live vector

10.2.3 Construction of Transfer Vector 188

10.2.4 Selection of insertion sites of foreign genes 188

10.3 poxvirus vector vaccine 188

Molecular biology of poxvirus 10.3. 189

10.3.2 vaccinia virus vector vaccine 190

10.3.3 avian pox virus vector vaccine 195

10.4 herpesvirus vector 209

10.4. 1 pseudorabies virus vector vaccine 209

10.4.2 1 bovine herpesvirus vector vaccine 2 17

10.4.3 Marek's disease virus vector 2 18

10.5 adenovirus 223

Overview of adenovirus vector 10.5. 1 223

Construction of 10.5.2 adenovirus vector 225

10.5.3 improved method of adenovirus vector 23 1

10.5.4 Prospect 234

10.6 retrovirus 235

10.6. 1 retrovirus overview 235

10.6.2 packaging principle of retrovirus vector 235

10.6.3 classification of retrovirus vectors 236

10.6.4 Construction of Retrovirus Vector 238

10.6.5 Advantages and disadvantages of retrovirus 238

10.6.6 Improvement of Retrovirus Vector in Recent Years 239

10.6.7 Prospect 240

10.7 A virus RNA replicon vaccine 24 1

10 . 7 . 38+04 1465437

Advantages and disadvantages of 10.7.2 A virus replicon vaccine 242

Immune mechanism of 10.7.3 A virus replicon vaccine 242

Several major replicators of viral RNA 243

Construction strategy of 10.7.5 A virus expression vector 246

Limitations of 10.8 recombinant live vector vaccine 247

10. 8. 1 safe247

10.8.2 maternal antibody interference 248

10.8.3 expression of recombinant virus 249

10.8.4 pathogenicity and immune efficacy 249

10.9 development direction of recombinant virus live vector vaccine 249

Reference 25 1

Chapter 1 1 bacterial vector vaccine 256

1 1. 1 salmonella carrier 256

Characteristics of Salmonella typhimurium strain11.1.12566.199689689686

Construction principle of attenuated Salmonella typhimurium 1 1. 1.2 strain. 36638.6666666666 1

1 1. 1.3 construction of a recombinant attenuated salmonella typhimurium vaccine and a case study on its immune mechanism50006.000000000505

1 1. 1.4 Construction method of recombinant Salmonella typhimurium strain 26 1

1 1. 1.5 conclusion 264

1 1.2 recombinant BCG vector 265

Advantages of 1 1.2. 1 recombinant BCG carrier.50000.000000000505

Research progress of 1 1.2.2 recombinant BCG carrier120066.1000006066666

1 1.2.3 the immune response induced by recombinant BCG vaccine and the influence of immune pathway on immune effect596661

Some problems in 1 1.2.4 RBCG 269

1 1.2.5 construction method of recombinant BCG vaccine5666/kloc-0 /2666.6666666668666

1 1.2.6 Outlook 273

1 1.3 Lactobacillus (Lactobacillus and Lactococcus lactis) vector 273

1 1.3. 1 lactobacillus carrier 274

1 1.3.2 Lactococcus 279

1 1.4 Vibrio vector vaccine 282

1 1.4. 1 Vibrio vector 282

1 1.4.2 is suitable for the expression system of attenuated Vibrio 283.

1 1.4.3 Maintenance of balanced lethal plasmid 283

1 1.4.4 More considerations about the application of bacterial vectors 284

1 1.5 Shigella vector vaccine 285

1 1.5. 1 Shigella carrier 285

1 1.5.2 Shigella transfer plasmid DNA285

1 1.6 listeria vector 285

Listeria vaccine 1 1.6. 1 286

1 1.6.2 listeria vector 286

1 1.6.3 the application of attenuated listeria in the treatment of tumor/kloc-0 /5666.6666666666666

1 1.6.4 listeria transfer plasmid DNA287

1 1.7 Bacillus subtilis integration vector 287

Research history of Bacillus 1 1.7. 1 integrated vector.50000.000000000605

1 1.7.2 the integration mechanism and integration mode of Bacillus subtilis integration carrier566661

1 1.7.3 construction of integrated vector vaccine against Bacillus subtilis120066.100000000666

1 1.7.4 Application of Bacillus Integration Vector 29 1

Reference 29 1

Chapter 12 nucleic acid vaccine 294

12.1the characteristics of DNA vaccine386661

Main advantages of 12. 1. 1 DNA vaccine50000.00000000016

12.1.2 Limitations of DNA vaccine 297

On the immune response mechanism of 12.2 DNA vaccine506661

6538+02. 2. 1 Talking about the DNA immune mechanism from the types of induced immune response56661

12.2.2 to explore the mechanism of DNA immunization from the aspect of inoculation route 30 1

Construction and evaluation of 12.3 DNA vaccine 303

Select the appropriate target gene and vector 36438. 6 1 56537.

Construction of 12. 3. 2 DNA vaccine 305

12.3.3 Improve the expression and immunogenicity of antigen protein 305

Verify the expression of antigen protein 306.

Evaluation of immune effect of 12.3.5 DNA vaccine 306

12.3.6 safety analysis

12.4 strategies to improve the immune effect of DNA vaccine 307

12.4. 1 Modification of the target gene encoding antigen 30077

Selection and Optimization of 12.4.2 Vaccine Plasmid Vector

12.4.3 gene adjuvant 309

12.4.4 ways and methods of immunization 3 15

12.4.5 improvement of DNA vaccine immunization method 3 17

Targeting strategy of 12.4.6 DNA vaccine

Vaccination plan 32 1

Improvement of 12.4.8 vaccine dosage form 322

Application and development trend of 12.5 DNA vaccine in disease control of livestock and poultry

12.5. 1 avian 323 related DNA vaccine

12.5.2 related DNA vaccine for pigs 325

12.5.3 bovine 326-related DNA vaccine

12.5.4 related DNA vaccine for dogs 327

12.5.5 DNA vaccine for bacterial diseases 327

Parasitic DNA vaccine 328

Safety of 12.6 DNA vaccine 329

Can the recombinant plasmid of 12.6. 1 be integrated into the genome of the host cell, which will lead to the activation of proto-oncogene or the inhibition of tumor suppressor gene? 30 109.00000000 1005

Can 12.6.2 recombinant plasmid induce autoimmune reaction and produce anti-DNA antibody 330?

Whether 12.6.3 recombinant plasmid continues to express foreign antigen will have adverse consequences.

12.6.4 Will the recombinant plasmid have reproductive toxicity 33 1?

12.6.5 Will the combination of recombinant plasmid and cytokines lead to other risks 33 1?

12.6.6 standardization of DNA vaccine

12.6.7 Risk-efficiency ratio 332

Conclusion 332

Reference 333

Reverse gene manipulation technology of influenza A virus and its application in vaccine development 56438+03 56686666 1

Study on the molecular biology of influenza virus 13. 1 10000.100000000806

Structure of influenza virus 13. 1. 1 and the function of its encoded protein110000.0000000000066

13. 1.2 influenza virus 339 replication

13.2 development of reverse genetics of influenza virus

13. 2. 1 Save the influenza virus with the help of helper virus 34 1

13.2.2 Cloning Gene to Save Influenza A Virus

On the application of 13.3 reverse gene manipulation technology in the research of influenza vaccine119666.10666666666666

13.3. 1 rescue of attenuated influenza vaccine strain/kloc-0 /40666.6066666666666

13.3.2 differential diagnosis of animal virus infection and vaccine immunity

13.4 experimental operation method of influenza virus 8 plasmid system 345

1 PHW 2000 bidirectional transcription vector 345

13.4.2 virus rescue with pHW2000 as the carrier 346

Potential customers 347

Reference 348

Chapter 14 reverse genetic manipulation of Newcastle disease virus and its application in the development of new vaccine 349

Reverse genetic manipulation of 14. 1 RNA virus30000.00000000105

14.1.1.349 Overview

Construction of 14. 1.2 infectious molecular clone

14.10.3 in vitro operation of infectious molecular cloning1666/6666666666666

Conclusion 353

Overview of Newcastle disease virus 14.2 353

Biological characteristics of Newcastle disease virus 14. 2+04120666/200666666666

14.2.2 molecular biological characteristics of Newcastle disease virus 357

14.2.3 Diagnosis, immunization and prevention of Newcastle disease 36 1

Principle and operation of reverse genetic operation of Newcastle disease virus 14.3

CDNA and other elements were constructed at the molecular level of 14.3. 1 36438 . 66666666667

14.3.2 rescue process 365

14.3.3 validation analysis 366

Matters needing attention in operation 367

Application of 14.4 reverse gene manipulation technology in Newcastle disease virus 367

14.4. 1 Study the relationship between structure and function 367

14.4.2 Recombinant virus 37 with reporter gene inserted1

14.4.3 labeled vaccine/chimeric vaccine 372

14.4.4 Poultry carrier vaccine 373

14.4.5 vaccine carrier for preventing human infectious diseases 374

14.4.6 vector modification for treating tumor 376

Conclusion 378

Reference 378

15 bacterial artificial chromosome technology and its application in the development of new vaccines Chapter 382

15. 1 bacterial artificial chromosome production technology 382

15.2 Structure, function and genetic characteristics of bacterial artificial chromosome 383

15.3 strategies and methods for constructing bacterial artificial chromosome 384

Construction strategy of bacterial artificial chromosome 15.3 366438+03636363636

15.3.2 Construction method of bacterial artificial chromosome 385

15.4 Construction and operation case of bacterial artificial chromosome 387

1 material 387

15.4.2 construction of recombinant Marek's disease virus transfer vector 387

15.4.3 Acquisition of Recombinant Marek's Disease Virus of Chicken 389

15.4.4 preparation of electrotransformation competent cell DH10b566436.00000000605

Screening of 15.4.5 BAC Molecular Clone Virus 389

15.4.6 BAC？ Determination of growth characteristics of recombinant virus rescued by DNA 390

Application and exploration of 15.5 bacterial artificial chromosome as vaccine 390

15.6 Modification System of Bacterial Artificial Chromosome Vaccine 393

15.6. 1RED/ET recombination system 93

Mechanism of 15. 6. 2 RED/ET recombination 394

15.7 Mechanism of Immune Response Induced by Bacterial Artificial Chromosome Vaccine 397

Advantages and disadvantages of 15.8 bacterial artificial chromosome vaccine

15.8. 1 Superiority of bacterial artificial chromosome vaccine506661

Problems and countermeasures of bacterial artificial chromosome vaccine 15.8.2/200661

15.9 prospect 398

Reference 399

Chapter 16 Joint immunization 400

16. 1 overview 400

The history and development of 16.2 combined vaccine 40 1

16.3 traditional combined vaccine 402

16.4 new combined vaccine 403

Manufacture and use of 16.5 combined vaccine 405

16.6 prospect 407

Reference 407

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