The first section the emergence of modern applied biotechnology 1
I traditional biotechnology 1
Second, modern biotechnology 2
Section 2 Contents of Modern Applied Biotechnology 2
Section III Global Development of Applied Biotechnology 4
The fourth quarter China application biotechnology development status 8
Section 5 Prospects of Modern Applied Biotechnology 12
Reference 13
Chapter II Genetic Engineering 14
Section 1 Overview 14
I. Gene research and its development 14
Second, the structure and function of DNA molecules 15
III. The Birth of Genetic Engineering 16
Four. Steps and contents of genetic engineering 17
Verb (abbreviation of verb) Safety of genetic engineering 18
Section 2 Various Tool Enzymes 18
I. Tool Enzymes and Genetic Engineering 18
Second, restriction endonuclease 19
3. Ligase 2 1
4. Modified enzyme 2 1
Section III Acquisition of Target Genes 23
I. Physical and chemical methods 23
Second, 24 shotguns
Three. Chemical synthesis of target gene 24
Four. Reverse transcription synthesis method 25
Verb (abbreviation for verb) polymerase chain reaction 25
Construction of genome library 26 of intransitive verbs
Seven. CDNA library 28
Section 4 Selection and Construction of Gene Vector 28
I. Gene vectors 28
Two. Plasmid cloning vector 29
Three. Viral and phage vectors 32
Four, chromosome positioning integration cloning vector 33
Verb (abbreviation of verb) artificial chromosome cloning vector 33
In vitro recombination and cloning of V region gene 34
I. Gene recombination 34
Second, the requirements of gene recombination on the vector 34
Three. Processing before connecting the target gene with the vector 35
Four, sticky end DNA fragment connection 36
Verb (abbreviation for verb) Connection of homologous terminal DNA fragments 37
Section 6 Gene Transfer and Recombinant Detection 38
I. introducing the recombinant into the recipient cell 39
Two. Screening and identification of recombinant clones 40
Section 7 Expression of Target Gene in Prokaryotes 42
I. Escherichia coli gene expression system 43
Second, the Bacillus expression system 45
Three. Isolation and purification of prokaryotic expression products 46
Section 8 Expression of Target Gene in Eukaryotic Cells 47
I. Yeast expression system 47
Second, the mammalian cell gene expression system 48
Reference 49
Chapter III protein Project 50
Section I Overview 50
I. Main Contents and Purpose of protein Project 50
Second, the foundation of protein engineering development 5 1
Three, protein engineering research methods on page 65
Section 2 Application of Fixed-point Mutation in protein Project 7 1
I. site-directed mutagenesis mediated by oligonucleotide 7 1
Second, the box mutation method 74
Thirdly, PCR induced mutation 74
Other methods of protein transformation in the third quarter 75
I. protein Modification 75
Second, the protein molecular design 78
The fourth quarter protein project progress 8 1
I. protein's structural forecast 82
Two, drug molecular design based on protein structure 84 pages.
Three, protein application engineering 85
Reference 87
The fourth chapter enzyme engineering 88
Section I Overview 88
I. Research and development of enzymes 88
Two. Introduction to enzyme engineering 89
Three. Overview of enzyme preparation industry at home and abroad 90 pages
Production of enzymes in the second quarter 92
I. Fermentation Process of Microbial Enzymes 93
Second, animal and plant raw material enzyme production and animal and plant cell culture enzyme production 96
3. Immobilized cells ferment to produce enzyme 97
Isolation and purification of the third enzyme 98
A, the general process of enzyme preparation 99
Two. Purification and Purification of Enzymes 100
Enzyme purity identification technology 104
Four. Preservation of enzymes 106
Section 4 Immobilization of Enzymes and Cells 106
I. Immobilization of enzyme 107
Second, cell immobilization 1 1 1
Three. Evaluation index of immobilized enzyme (cell) 1 12
Four. Application of Immobilized Enzyme (Cell) 1 12
Section 5 Enzyme Reactor 1 15
I. Types and characteristics of enzyme reactors 1 15
Second, the design and selection of enzyme reactor 1 17
Section VI Other Research Fields of Enzyme Engineering 1 18
I. Chemical Modification of Enzymes
2. Enzymatic reaction in non-aqueous medium 120
Iii. Artificial mimic enzyme 122
IV. Ribozymes and Deoxyribozymes 125
V directed evolution of enzymes 126
Section 7 Application of Enzymes 128
I application of enzymes in food industry 128
Second, the application of enzymes in light industry 132
Iii. Application of Enzymes in Medicine 134
Four. Application of enzyme in analysis and detection 136
Application of verb (abbreviation of verb) enzyme in environmental protection 137
Reference 138
Chapter 5 Monoclonal Antibodies and Antibody Engineering 139
Section 1 antibody overview 139
Introduction to antibody development 139
Second, the structure and function of antibody 140
Section 2 Generation of Monoclonal Antibodies 14 1
I introduction 14 1
Second, the production of monoclonal antibodies 14 1
In the third quarter, genetically engineered antibody 145
1. Humanization of mouse monoclonal antibody 145
Second, the single chain antibody 146
Three. Antibody fusion protein 146
Four. Phage antibody 149
Section 4 Application of Antibodies 150
I. Application of Antibodies in the Treatment of Clinical Diseases 150
Second, the application of antibodies in disease diagnosis 156
Section 5 Expression System of Antibody 159
I mammalian cell expression system 159
2. Escherichia coli expression system 159
Three. Yeast expression system 160
Iv. insect cell expression system 160
Verb (abbreviation of verb) animal and plant expression system 160
Reference 16 1
Chapter VI Modern Molecular Diagnostic Technology 162
Section 1 Enzyme-linked Immunosorbent Assay 163
I. Principle of Enzyme-linked Immunosorbent Assay 163
Second, enzyme-linked immunosorbent assay reagents and materials 163
III. Commonly Used Enzyme-linked Immunodiagnostic Techniques 164
Four. Selection of the best working concentration of enzyme-linked immunosorbent assay
Application of verb (abbreviation of verb) ELISA 169
Limitations of enzyme-linked immunosorbent assay 170
Section 2 DNA diagnostic technology 17 1
I. nucleic acid hybridization 17 1
Molecular diagnosis of malaria 184
III. Molecular Diagnostic Techniques for Bacterial Infectious Diseases and Viral Diseases 184
Section III Molecular Diagnosis of Genetic Diseases 187
I. Genetic Disease Detection System 187
Second, the selection of genetic diagnosis for genetic diseases 19 1.
Third, the application of PCR technology in the diagnosis of genetic diseases 193
Section IV Molecular Diagnostic Techniques for Cancer 194
I. Oncogenes and tumor suppressor genes 195
Ii. Gene changes in tumorigenesis 197
Three. Detection and evaluation of proto-oncogene and tumor suppressor gene 199
Section 5 Detection of Environmental Microbes 20 1
First, the application of PCR technology in the detection of environmental microorganisms 20 1
Reference 204
Chapter VII Modern Biotechnology and Vaccine 206
Section 1 Overview 206
I. research and development of vaccines 206
Two. Vaccine type 207
Three. Application of vaccine
The application of modern biotechnology in vaccinology +0 1
Molecular biology and vaccinology 2 1 1
Ii. technical research on vaccines 2 14
Section 3 Genetic Engineering Vaccine 2 17
I. Introduction to 2 17
Immunological basis of genetically engineered vaccine 2 17
Three. Reactivity and immunogenicity of genetically engineered vaccine 2 18
IV. Research Progress of Genetically Engineered Vaccines 2 18
Verb (abbreviation of verb) Development of genetically engineered hepatitis B subunit vaccine 2 19
Six, the application prospect of genetic engineering vaccine 220
The fourth quarter nucleic acid vaccine 220
First, the theoretical basis of nucleic acid vaccine 220
Second, the mechanism of action of nucleic acid vaccine 222
Third, the construction method of nucleic acid vaccine 222
Four. Methods and approaches of DNA vaccine immunization 225
Development status and trend of verb (abbreviation of verb) DNA vaccine 226
Section 5 Recombinant Live Vaccines and Their Vectors 227
I. Living recombination vaccines 227
Second, the live vaccine vector 228
Three. Menger virus vector 230
Four, canary pox virus vector 230
Verb (abbreviation of verb) Sendai virus vaccine vector 23 1
Six, measles virus vaccine vector 23 1
Seven, Salmonella vaccine carrier 23 1
Section 6 Bacterial vaccines and other vaccines 232
I. Antimicrobial vaccine 232
Two. Antiidiotypic antibody vaccine 233
Three. Plasmodium falciparum vaccine 233
Four. Contraceptive vaccine 234
Section 7 Tumor Vaccine 235
I. Tumor cell vaccine 235
Second, the embryo antigen vaccine 236
Three. Virus vaccine 236
Four. Cancer vaccine 236
Verb (abbreviation of verb) synthetic peptide tumor vaccine 237
Intransitive verb anti-idiotypic vaccine 237
7. Preparation of tumor vaccine 238 with tumor cell shock APC.
Eight, using bispecific antibody to modify tumor cells to prepare tumor vaccine 238.
Section 8 AIDS vaccine 238
I. Structure and infection characteristics of HIV 239
Two. HIV genome and coded protein 239
Three. Gene expression after HIV invades human body 240
Four. Animal model of experimental AIDS vaccine 240
Verb (abbreviation of verb) Research status of AIDS vaccine 24 1
Section 9 autoimmune disease vaccine 242
Section 10 Therapeutic vaccine 244
A, viral infectious disease treatment vaccine 244
II. Vaccines for cardiovascular diseases 244
Three, the treatment of blood infection caused by bacteria vaccine 245
Four, other applications 245
Verb (abbreviation of verb) Appendix: Milestone of Immune Development 245
Reference 246
Chapter VIII Biotechnology and Pharmacy 248
The first section natural medicine pharmacy 248
I. Drugs of plant origin 248
Two. Animal-derived drugs 252
Three. Human drug 255
Four. Natural marine medicine 256
V. Microbes and mineral drugs 258
In the second quarter, genetic engineering drugs 258
I. Overview 258
Two. Classification of genetically engineered drugs 259
Three, the production of genetically engineered drugs 260
Four, genetic engineering drug preparation example 263
Reference 265
Chapter 9 Gene Therapy 267
The first section overview 267
I. Research and development of gene therapy 267
Second, the current situation of gene therapy 268
The second part of the gene transfer vector 270
I. Introduction 270
Second, retrovirus vector 27 1
Three. Adenovirus vector 27 1
Four. Adeno-associated virus vector 272
Verb (abbreviation of verb) herpes simplex virus vector 272
Intransitive verb vaccinia virus vector 273
Seven. Non-viral vector 274
In the third quarter, the way of gene therapy 276
First, the strategy of gene therapy 276
Second, the ways and methods of gene transfer in gene therapy 277
Three. Target tissue of gene therapy 278
The fourth quarter gene therapy for common diseases 278
A, blood circulation disease gene therapy 278
Second, the gene therapy of cardiovascular diseases 279
Three. Gene therapy of tumor 28 1
Four, suicide gene therapy 283
Verb (abbreviation of verb) Gene therapy for HIV infection 285
Six, gene therapy for other diseases 286
Seven. The prospect of gene therapy
Reference 288
Chapter 10 Modern Fermentation Engineering 289
The first section overview 289
Brief history of fermentation industry development 289
Second, the characteristics of the fermentation industry 292
Three. Application scope of fermentation industry 293
Fourth, the significance and prospect of modern fermentation engineering 296
In the second quarter microbial fermentation 296
A, commonly used in industrial production strains and microorganisms 296
Second, the culture medium 299
Three. Disinfection 304
Four, the general process of fermentation 306
Section III Breeding and Preservation of Strains
I. Natural reproduction 308
Second, mutation breeding 309
III. Hybrid Breeding 3 14
Four, metabolic control breeding 3 18
V. Genetic Engineering Breeding 3 19
VI. Preservation and rejuvenation of microbial strains 32 1
The fourth quarter microbial metabolism and regulation 324
I. Metabolism of Microorganisms 324
Second, the regulation of microbial primary metabolism 325
Third, the regulation of microbial secondary metabolism 327
Section 5 Fermentation Process Control 33 1
1. The main control parameter in the fermentation process is 33 1.
Second, the metabolic changes in the fermentation process 333
III. Influence of Substrate and Control of Feeding 334
Four, the influence of temperature and its control 337
The influence of verb (verb's abbreviation) pH and its control 339
Six, the influence of dissolved oxygen and its control 340
Seven, CO? 2 and the influence of its control 342
Eight, the influence of the bubble and its control 344
Nine, the determination of fermentation endpoint 346
X. control of fermenting bacteria
XI。 Fermentation control of genetically engineered strain 350
Section 6 Extraction and Refining of Fermented Products 353
I. Pretreatment and filtration of fermentation broth 354
Second, the extraction of fermented products 355
Three. Refinement of fermented products 357
Reference document 360
Chapter 11 Biotechnology and Food 362
The first section biotechnology and food processing 362
I. Biotechnology and protein Food 362
II. Biotechnology and Brewed Food 365
Three. Biotechnology and beverage industry 368
Four, food additives 372
Verb (abbreviation of verb) functional food 374
In the second quarter biotechnology and food testing 376
A, the application of immunological technology in food testing 376
Second, the application of molecular biology technology
In the third quarter genetically modified food 378
I. Overview of Genetically Modified Foods 378
Second, the detection of genetically modified food 380
III. Safety and Prospect of Genetically Modified Foods 382
Reference 383
Chapter XII Modern Biotechnology and Energy Development
The first section microbial metallurgy 384
A, the principle of microbial metallurgy 384
Two. Microorganisms used in metallurgical industry
Three. Industrial method of bacterial leaching 387
Four, the practical application of microbial metallurgy technology 388
Verb (abbreviation of verb) Prospect of microbial metallurgy
In the second quarter fuel ethanol 390
First, the significance of using fuel ethanol 390
Second, the production of fuel ethanol 39 1
Three, ethanol gasoline blending method 392
Fourth, the development of fuel ethanol 393
In the third quarter biogas fermentation 394
I. Significance of developing biogas 394
Second, biogas fermentation 395
Reference 398
Chapter 13 Modern Biotechnology and Environmental Protection 400 pages
Section 1 Microbial Purification of Sewage 400
I. Overview of Biological Treatment of Wastewater 400
Second, aerobic biological treatment technology 404
Three. Anaerobic biological treatment process 408
In the second quarter, atmospheric purification biotechnology 4 10
First, the principle of waste gas biological treatment 4 1 1
Second, the waste gas biological treatment process 4 1 1
Three. Biological treatment of carbon dioxide 4 13
Four. Present situation and prospect of waste gas biological treatment 4 15
Section 3 Solid Waste Disposal 4 16
I. Composting Method 4 16
II. Landfill Technology 4 18
Section IV Bioremediation Technology 4 18
I. Basic principles and main methods of bioremediation 4 18
Second, the bioremediation technology of soil pollution 420
Three. Bioremediation technology of groundwater pollution
Section 5 Genetic Engineering and Pollution Control 424
1. Genetically engineered bacteria for degrading halogenated aromatic hydrocarbons 425
Second, genetically engineered bacteria that degrade herbicides 425
Three, genetically engineered bacteria to control pesticides 425
Four, petroleum degradation functional bacteria 425
Section VI Biotechnology for Environmental Pollution Monitoring and Assessment
I. biological monitoring and assessment of environmental pollution 426
Second, the application of nucleic acid probe in environmental monitoring 430
Iii. PCR technology 43 1
Four. Biosensor 43 1
Reference 435
Chapter XIV Agricultural Biotechnology and Its Application
The first section plant cell engineering 436
First, plant tissue culture
Second, plant protoplast culture and somatic hybridization technology 443
In the second quarter plant transgenic technology and application 446
I. Plant Gene Transformation Technology 447
Two, the application of transgenic plants 450 pages
In the third quarter animal cloning technology 458
I. Concept and significance of animal cloning 458
Second, the animal embryo segmentation technology 460
Three, animal embryo nuclear transfer technology 46 1
Four, animal differentiated nuclear transfer 463
Five, the main technical operation examples of animal cloning 463
VI. Development and industrialization of animal cloning technology 464
VII. Problems of cloning technology 466
The fourth quarter transgenic animal research 466
I. Overview of Transgenic Animals Research
Second, the animal transgenic technology 468
III. Genetically modified livestock, fish and poultry 47 1
Four. Problems and prospects of animal transgenic 474
Reference 474
Chapter 15 Biological pesticides 476
Section 1 Overview 476
I. Classification of biological pesticides 476
Two. Source, Mechanism and Target of Biological Pesticide 477
The toxicity and efficacy of biological pesticides in the second quarter 482
I. Concepts of toxicity and efficacy 483
Two. Expression and analysis of virulence 483
III. Expression and analysis of efficacy 485
Section III Development, Production and Use of Biological Pesticides
I. Development and production of bacterial insecticides 488
Second, the development and production of fungal pesticides 490
Third, the development and production of virus pesticides 493
Four. Development and production of nematodes and protozoan insecticides
Development and production of verb (abbreviation of verb) agricultural antibiotic 495
Development and production of intransitive verb microbial herbicide 495
Seven, the development and production of plant-derived biochemical pesticides 496
Eight, biological pesticide processing dosage forms and additives 496
Nine, the use of biological pesticides 497
The fourth quarter biological pesticide quality inspection 498
I. Beauveria bassiana 498
Second, Bacillus thuringiensis 499
Three. Insect virus insecticide 499
Four, agricultural antibiotics 500
Section 5 Development and application prospect of biological pesticides 50 1
Reference document 503
Chapter 16 Microbial fertilizer 504
Section 1 Overview 504
I. Meaning and characteristics of microbial fertilizer 504
Second, the main types of microbial fertilizer 505
Third, the role of microbial fertilizer 506
In the second quarter, the plant nutrient cycle and its function 508
I. Nitrogen Cycle 508
Two. Phosphorus cycle 5 1 1
Three. Potassium cycle 5 12
Iv. Mineral elements necessary for plants and their functions 5 13
Verb (abbreviation of verb) Characteristics of mineral elements absorbed by plant roots 5 18
Section 3 Nitrogen Fixing Bacteria Fertilizer and Rhizobium Fertilizer 5 19
I biological nitrogen fixation 5 19
Second, nitrogen-fixing bacteria fertilizer 525
III. Rhizobium Fertilizer 528
The fourth quarter phosphate-solubilizing microbial fertilizer and silicate bacteria fertilizer 530
I. Phosphorus-solubilizing microbial fertilizer 530
Second, silicate bacterial fertilizer 53 1
Section 5 Microbial fertilizer for rhizosphere growth-promoting bacteria 532
I. Species of rhizosphere growth-promoting bacteria 532
Second, the mechanism of rhizosphere growth-promoting bacteria preparation 532
III. Attribution of Rhizosphere Growth-promoting Bacterial Preparations 533
Four. Application status of rhizosphere growth-promoting microbial inoculum 533
Research and application prospect of verb (abbreviation of verb) rhizosphere growth-promoting bacteria preparation 534
Section 6 Other Microbial Fertilizers 534
I. Microbial fertilizer for decomposing crop straws 534
Second, the compound microbial fertilizer 535
Three. AM mycorrhizal microbial fertilizer 535
Four, 5406 antibiotic fertilizer 536
Section 7 Development Strategy of Microbial Fertilizer in China 536
First, the development status and existing problems of microbial fertilizer in China
Second, the development trend and development strategy of microbial fertilizer in China 538
Three, China microbial fertilizer industrialization development prospects 538
Section 8 Requirements for Registration Data of Microbial Fertilizer 539
I. Summary of fertilizer registration data 539
Second, the fertilizer registration information 540
Three. Related accessories 545
Reference 548
Chapter 17 Human Genome Project and Post-genome Research 549
Section 1 Overview of the Human Genome Project 549
I. Historical and technical background of the emergence of the Human Genome Project 549
Second, the task and significance of the human genome project 553
Three. Main contents of human genome research
Four, the strategy of human genome research 555
V. Chronology of Human Genome Project 555
In the second quarter, the influence of human genome project on medical development 555
A new era of genetic medicine, 555
Second, the development of gene drugs 558
In the third quarter, the protection of gene resources 559
I. Competition for genetic resources 559
II. Protection of Genetic Resources in China 56 1
Research progress of genomics after the fourth quarter 563
I. Post-genomic Era 563
Second, the research progress of post-genome 565
Reference 568
Chapter XVIII Application Rules and Invention Protection of Modern Biotechnology
Section 1 Overview 570
Section 2 Quality Management Standard for Non-clinical Research of Biotechnology Drugs (GLP) 57 1
I. Concept of GLP 57 1
Second, the development of GLP 57 1
Three. The product range is governed by GLP in various countries.
Four. Bilateral agreements and international cooperation of GLP 573
Use of verb (abbreviation of verb) GLP formula 573
The basic content of intransitive verb GLP 574
Seven, the main problems of GLP laboratory identification 578
Section III Clinical Trial of GMP (GCP) 579 Biotechnology Drugs
First, the concept of GCP
Second, the development of GCP 580
Three. Application of GCP formula 58 1
Four. International integration of GCP 58 1
Verb (abbreviation of verb) The important role of GCP 582
Basic principles of intransitive verb GCP 582
Seven, the basic content of GCP 583
Section 4 Good Manufacturing Practices for Biotechnology (GMP) 584.
First, the concept of GMP 584
Second, the development of GMP 584
Iii. Introduction to GMP 585
Section 5 Good Quality Management Practice for Biotechnology Drugs (GSP) 59 1
I. Concept of GSP 59 1
Second, the development of GSP 592
III. Purpose of GSP formulation 592
Four. Main contents of GSP 592
Section 6 Patent Protection of Modern Biotechnology
I. Introduction 595
II. Patent Protection for Biotechnology Inventions and Innovations 596
Three, the characteristics of biotechnology invention and innovation patent 596
Four, modern biotechnology patent type 597
Verb (abbreviation of verb) The urgency and influence of patent protection for biotechnology inventions.
Reference 599
Chapter 19 Safety and Social Ethics of Modern Biotechnology 600
Section 1 Introduction 600
I. Debate on Biosafety in the World 600
Two. Understanding and management of biosafety in the world 60 1
Three. Biosafety assessment
In the second quarter, the safety of modern biotechnology 607
I. Safety of genetically modified crops 607
Second, the safety of transgenic animals 6 10
Third, the safety of genetically modified foods 6 1 1
Four. Biological Weapons 6 12
Section 3 Social Ethics of Modern Biotechnology 6 14
I. Human Genome Project and Social Ethics 6 14
Ii. Social and Ethical Issues Caused by Animal Cloning and Human Cloning 6 15