Infodoc : Sciences, Techniques et Culture

Part 1: Application and Perspectives
1 Transgenic Modification of Production Traits in Farm Animals
1.1 The Creation of Transgenic AnimaIs
1.1.1 Pronuclear DNA Microinjection
1.1.2 Retroviral Vectors
1.1.3 Pluripotent Stem Cell Technologies
1.1.4 Nuclear Transfer using Transgenic Cells
1.1.5 Gene Transfer in Poultry
1.1.6 Gene Transfer in Fish
1.2 Transgenes: Gene Constructs
1.3 Transgenic AnimaIs with Agricultural Traits[...]

Part 1: Application and Perspectives
1 Transgenic Modification of Production Traits in Farm Animals
1.1 The Creation of Transgenic AnimaIs
1.1.1 Pronuclear DNA Microinjection
1.1.2 Retroviral Vectors
1.1.3 Pluripotent Stem Cell Technologies
1.1.4 Nuclear Transfer using Transgenic Cells
1.1.5 Gene Transfer in Poultry
1.1.6 Gene Transfer in Fish
1.2 Transgenes: Gene Constructs
1.3 Transgenic AnimaIs with Agricultural Traits
1.3.1 Improved Growth Rate, Carcass Composition, and Feed Efficiency
1.3.1.1 Transgenes in mammalian farm animaIs
1.3.1.2 Transgenes in fish
1.3.2 Alteration of Milk Composition
1.3.3 Improved Animal Health
1.3.3.1 Additive gene transfer of resistance genes
1.3.3.2 Gene targeting of susceptibility genes
1.3.4 Improved Biochemical pathways
1.3.5 Improved Wool Production
1.4 Transgenic Farm AnimaIs and Biosafety Issues
1.5 Conclusions
2 Genetically Modified Plants
2.1 Methods to Establish Genetically Modified Plants
2.1.1 Transformation Methods
2.1.1.1 Agrobacterium transformation
2.1.1.2 Direct gene transfer
2.1.2 Tissue Requirements
2.1.3 Molecular Requirements
2.1.3.1 Promoter
2.1.3.2 Codon usage
2.1.3.3 Selectable marker and reporter genes
2.2 GM Plants Already on the Market (EU, USA, Canada, Japan)
2.2.1 Herbicide Resistance in Soybean, Maize, Oilseed rape, Sugar beet, Rice, and Cotton
2.2.2 Insect Resistance in Maize, Potatoes, Tomatoes, and Cotton
2.2.3 Virus-resistance, male sterility, delayed fruit ripening, and fatty acid contents in GMPs
2.3 GM Plants "in the Pipeline"
2.3.1 Input Traits
2.3.1.1 Insect resistance in rice, soybean, oilseed rape, eggplant, walnut, grape, and peanut
2.3.1.2 Disease resistance in maize, potatoes, fruits, and vegetables
2.3.1.3 Tolerance against abiotic stresses
2.3.1.4 Improved agronomic properties
2.3.2 Traits Affecting Food Quality for Human Nutrition
2.3.2.1 Increased carotenoid content in rice and tomato
2.3.2.2 Elevated iron level in rice and wheat
2.3.2.3 Improved amino acid composition in potato plants
2.3.2.4 Reduction in the content of antinutritive factors in cassava
2.3.2.5 Production of "low-calorie sugar" in sugar beet
2.3.2.6 Seedless fruits and vegetables
2.3.3 Traits that Affect Processing
2.3.3.1 Altered gluten level in wheat to change baking quality
2.3.3.2 Altered grain composition in barley to improve malting quality
2.3.4 Traits of Pharmaceutical Interest
2.3.4.1 Production of vaccines
2.3.4.2 Production of pharmaceuticals
2.4 Outlook
3 Fermented Food Production using Genetically Modified Yeast and Filamentous Fungi
3.1 Introduction
3.1.1 Why Do We Ferment Foodstuffs?
3.1.2 Fermented Foods of Plant Origin
3.1.3 Fermented Foods of Animal Origin
3.1.4 Conclusion
3.2 Application of Recombinant DNA Methods
3.2.1 Recombinant DNA Technology in Yeast
3.2.1.1 Vectors
3.2.2 Recombinant DNA Technology in Filamentous Fungi
3.2.2.1 Fungal transformation
3.3 Improved Fermentation Efficiency for Industrial Application
3.3.1 Industrial Saccharomyces Strains
3.3.1.1 Beer
3.3.1.2 Wine
3.3.1.3 Sake
3.3.1.4 Bread
3.3.2 Other Industrial Yeast Strains
3.3.3 Industrial Filamentous Fungi
3.4 Commercial use of Genetically Modified Organisms (GMO)
3.5 The Future
4 Production of Food Additives using Filamentous Fungi
4.1 Filamentous Fungi in Food Production
4.1.1 Industrial Applications
4.2 Additives for the Food Industry
4.3 Design of GMM for Production of Food Additives and Processing Aids
4.3.1 Gene Disruption
4.3.2 Expression Vectors
4.4 Industrial Enzyme Production Processes
5 Perspectives of Genetic Engineering of Bacteria used in Food Fermentations
5.1 Introduction
5.2 Lactic Acid Bacteria
5.2.1 Lactococcus lactis subsp. lactis and subsp. cremoris
5.2.2 Lactobacillus spp.
5.2.3 Streptococcus thermophilus
5.2.4 Leuconostoc spp.
5.2.5 Pediococcus spp.
5.2.6 Oenococcus spp.
5.3 Perspective and Aims
5.3.1 Bioconservation
5.3.2 Bacteriophage Resistance
5.3.3 Exopolysaccharides
5.3.4 Proteolysis
5.4 Metabolic Engineering of Lactic Acid Bacteria
5.5 Stress Responses in Lactic Acid Bacteria
5.6 Methods
5.6.1 Transformation and Vector Systems
5.7 Conclusions
Part Il: Legislation in Europe
6 The Legal Situation for Genetically Engineered Food in Europe
6.1 Introduction
6.1.1 The Need for Regulation
6.1.2 The History of the Novel Food Regulation
6.2 Status Quo
6.2.1 The Novel Food Regulation
6.2.1.1 Introduction
6.2.1.2 Scope of application
6.2.1.3 Requirements for novel foods and food ingredients
6.2.1.4 Procedures
6.2.1.5 Labeling
6.2.1.6 Other questions
6.2.2 Problems
6.2.2.1 Negative labeling
6.2.2.2 Compliance with World Trade law
6.2.2.3 Competent authorities in the member states
6.2.4 Supplementary and Replacement Regulation
6.2.4.1 History
6.2.4.2 Scope of application of the Replacement Regulation
6.2.4.3 Requirements of labeling
6.2.5 Relation to Council Directive No 90/220/EEC of 23 April 1990 on the deliberate release into the environment of genetically modified organisms (OJ (EC) 1990 No L 117/15; from here on: Deliberate Release Directive)
6.2.6 Supplementary National Provisions in German law: The Novel Foods and Food Ingredients Instrument
6.2.6.1 General rules for novel foods
6.2.6.2 Rules on labeling of genetically modified soya beans and genetically modified maize
6.2.6.3 Availability of negative labeling of foodstuffs made without using procedures of genetic engineering
6.2.6.4 Rules on criminal offences and misdemeanors
6.3 Recent Development at the European Level
6.3.1 Introduction
6.3.2 The Commission ProposaI for a "Regulation of the European Parliament and of the Council on Genetically Modified Food and Feed"
6.3.2.1 Objective and definitions
6.3.2.2 Requirements of genetically modified food and feed
6.2.2.3 Labeling of genetically modified food and feed
6.3.2.4 General provisions
6.3.3 The Commission ProposaI for a Regulation of the European Parliament and of the Council Concerning Traceability and Labeling of Genetically Modified Organisms and Traceability of Food and Feed Products Produced from Genetically Modified Organisms and Amending Directive 2001/18/EC
6.3.4 Stage of Legislative Procedure
Part III: Methods of Detection
7 Detection of Genetic Modifications: Some Basic Considerations
7.1 The Conversion of Genetic Information from DNA to Phenotypes
7.2 DNA, Protein and Phenotypes as Targets for Detection Assays
7.3 Food-grade Modifications
7.4 Detection of Unknown Modifications
8 DNA-based Methods for Detection of Genetic Modifications
8.1 Introduction
8.2 Recent DNA Methodology
8.2.1 Sampling Procedure
8.2.2 Extraction and Purification of DNA
8.3 Specific Detection of Genetic Material
8.3.1 DNA Hybridization-based Detection Technique (Southern BIot)
8.4 Nucleic Acid Amplification Methods using PCR
8.4.1 The Common PCR
8.4.2 Real-time PCR
8.4.3 Important Bioinformatic Considerations
8.5 Alternative and Promising DNA Detection Techniques
8.5.1 Thermal Cycling Procedures
8.5.2 Isothermic Amplification
8.5.3 DNA-micro-arrays
8.5.4 Mass Spectrometry (MS) of DNA
8.5.5 Supplementary Photon-driven Monitoring Methodologies
8.5.6 Novel Biological Monitoring Approaches
8.6 Conclusions and Future Prospects of GMO Detection Applying DNA-analysis
9 Genetic Engineering of Fishes and Methods for Detection
9.1 Introduction
9.2 Development and Production ofTransgenic Fish 9.2.1 Structure of Gene Cassettes
9.2.2 Methods of Gene Transfer
9.2.3 Evidence for Gene Transfer and Expression
9.3 Examples of Successful Production of Transgenic Fish
9.3.1 Atlantic Salmon
9.3.2 Pacific Salmon
9.3.3 Tilapia (O. hornorum hybrid)
9.3.4 Tilapia (O. niloticum)
9.3.5 Carp
9.4 Methods to Detect Processed Transgenic Fish
9.5 Food Safety of Transgenic Fish
9.5.1 The Gene Product
9.5.2 Pleiotropic Effects
10 Detection Methods for Genetically Modified Crops
10.1 Introduction
10.2 Isolation of Plant DNA
10.2.1 Sampling
10.2.2 Sample Preparation
10.2.3 DNA Extraction and Analysis
10.3 Detection Strategies
10.3.1 Screening
10.3.2 Specific Detection
10.3.2.1 Example for Qualitative Detection
10.3.3 Quantification
10.3.4 Verification
10.3.5 Validation
10.4 Outlook and Conclusions
11 Methods to Detect the Application of Genetic Engineering in Composed and Processed Foods
11.1 Introduction
11.2 Challenges Specific to the Detection of GMO in Composed and Processed Foods
11.3 Degradation of Proteins and DNA
11.3.1 Proteins
11.3.2 DNA
11.4 Analytical Approaches
11.4.1 Protein-based Methods
11.4.2 DNA-based Methods
11.4.2.1 Qualitative PCR
11.4.2.2 Quantitative PCR
11.4.2.3 Competitive PCR
11.4.2.4 Real-time PCR
11.5 Conclusions
12 Mutations in Lactococcus lactis, and their Detection
12.1 Introduction
12.2 Composition of the Genome of Lactococcus lactis
12.3 Flexibility in the Genome of Lactococcus lactis
12.3.1 Conjugation
12.3.2 Transduction
12.3.3 Transformation
12.3.4 IS Elements and Transposons
12.3.5 Lactococcal Phage as Sources of Genetic Plasticity
12.3.5.1 An example of natural genetic flexibility: the Lactococcus lactis NCDO712 family
12.4 Mutations in Lactococcus lactis as a Conseqequence of Environmental Factors and DNA Metabolism
12.5 Methods to Mutate the Genome of Lactococcus lactis
12.5.1 Genetic Engineering of Lactococcus lactis
12.6 Strategies to Detect Genetically Modified Lactococcus lactis
12.6.1 Sample Preparation
12.6.2 DNA-based Procedures
12.6.2.1 Southern hybridization
12.6.2.2 PCR
12.6.3 Nucleotide Sequence-based Procedures
12.6.3.1 Micro-arrays
12.6.4 Protein-based Procedures
12.6.4.1 Western hybridization
12.6.4.2 ELISA
12.6.4.3 SPR
12.6.4.4 Two-dimensional gel electrophoresis and mass spectrometry
12.7 Conclusions
13 Detection Methods for Genetically Modified Microorganisms used in Food Fermentation Processes
13.1 Introduction
13.2 Properties of Microorganisms
13.3 Current Methods for Detection of GMM
13.3.1 DNA Isolation
13.3.2 DNA Stability
13.3.3 Organism-specific Detection of GMM
13.4 Conclusion
Index