| Selectable and Screenable Markers for Rice Transformation | |
| Introduction | p. 1 |
| Dominant Selectable Markers for Rice | p. 3 |
| Aminoglycoside 3'-Phosphotransferase (Neomycin Phosphotransferase) | p. 3 |
| Hygromycin Phosphotransferase | p. 4 |
| Phosphinothricin Acetyltransferase | p. 4 |
| Other Dominant Selectable Markers | p. 5 |
| Novel Selectable Markers | p. 7 |
| Innocuous Markers | p. 7 |
| Counterselectable Markers | p. 7 |
| Screenable Marker Genes | p. 8 |
| [beta]-Glucuronidase (gusA) | p. 9 |
| Firefly Luciferase (luc) | p. 11 |
| Green Fluorescent Protein (gfp) | p. 11 |
| Strategies for Marker-Gene Delivery | p. 13 |
| References | p. 14 |
| Use of Green Fluorescent Protein to Detect Transformed Shoots | |
| Introduction | p. 19 |
| GFP: Suitable as a Visually Selectable Marker In Planta? | p. 20 |
| Important Properties of the Protein | p. 20 |
| Properties of a Useful Selectable Marker in Plant Transformation Technology | p. 21 |
| GFP Expression and Detection in Primary Transformed Tissues | p. 22 |
| Transient Expression and GFP Detection | p. 22 |
| Detection Equipment and Troubleshooting | p. 22 |
| Stable Expression and GFP Detection in Primary Transformed Tissues | p. 23 |
| GFP for Screening of Segregating Populations | p. 28 |
| Conclusion | p. 29 |
| References | p. 29 |
| Luciferase Gene Expressed in Plants, Not in Agrobacterium | |
| Introduction | p. 31 |
| Preventing Bacterial Expression | p. 31 |
| Imaging Luciferase Activity In Planta | p. 32 |
| Measuring Luciferase Activity in Plant Extracts | p. 34 |
| References | p. 35 |
| Use of [beta]-Glucuronidase (GUS) To Show Dehydration and High-Salt Gene Expression | |
| Introduction | p. 37 |
| What Is GUS? | p. 39 |
| Trasgenic Plants Carrying Promoter-GUS Constructs | p. 41 |
| Construction of Promoter-GUS Fusion Genes | p. 44 |
| Introduction of Promoter-GUS Constructs into Agrobacterium | p. 45 |
| Transformation of Plants with Agrobacterium | p. 46 |
| Transformation of Arabidopsis Plants | p. 46 |
| Fluorometric Assay | p. 49 |
| Introduction | p. 49 |
| Stress Conditions | p. 49 |
| Plant Preparation | p. 49 |
| Dehydration | p. 49 |
| High Salinity | p. 49 |
| ABA Treatment | p. 50 |
| Other Treatments | p. 50 |
| Protein Assay | p. 50 |
| Sample Preparation | p. 50 |
| Fluorometric Assay | p. 51 |
| Histochemistry | p. 51 |
| Introduction | p. 51 |
| Histochemistry | p. 52 |
| Northern Analysis of GUS | p. 52 |
| Introduction | p. 52 |
| RNA Extraction | p. 53 |
| RNA Blotting | p. 54 |
| Northern Hybridization | p. 55 |
| Application of the GUS System | p. 56 |
| Transient Assay | p. 56 |
| Transactivation Experiment | p. 57 |
| Promoter Tagging (Enhancer Trap) | p. 57 |
| Conclusion | p. 57 |
| References | p. 59 |
| Methods for Detecting Genetic Manipulation in Grain Legumes | |
| Introduction | p. 63 |
| Detection at the DNA Level | p. 64 |
| PCR Analysis | p. 65 |
| Control PCR and Specific PCR Systems | p. 66 |
| Quantitative Approach | p. 69 |
| Competitive PCR | p. 69 |
| Real-Time PCR Systems | p. 70 |
| Concluding Remarks | p. 70 |
| References | p. 71 |
| Elimination of Selectable Marker Genes from Transgenic Crops | |
| Introduction | p. 73 |
| Co-transformation | p. 74 |
| Transposon-Mediated Approaches | p. 78 |
| Transposon-Mediated Repositioning | p. 78 |
| Transposon-Mediated Elimination | p. 80 |
| Site-Specific Recombination | p. 81 |
| The Cre/loxP System | p. 82 |
| The FLP/frt System | p. 86 |
| The R/RS System | p. 88 |
| Intrachromosomal Homologous Recombination | p. 89 |
| Conclusions and Future Prospects | p. 90 |
| References | p. 91 |
| GST-MAT Vector for the Efficient and Practical Removal of Marker Genes from Transgenic Plants | |
| Introduction | p. 95 |
| ipt-Type MAT Vectors | p. 96 |
| Transposable Element | p. 96 |
| Site-Specific Recombination System | p. 97 |
| Advantages of the ipt Gene | p. 101 |
| Two-Step Transformation | p. 103 |
| Promoter of the R Gene | p. 103 |
| Promoter of the ipt Gene | p. 105 |
| Combination of the ipt and iaaM/H Genes | p. 108 |
| Transgene Stacking | p. 110 |
| Single-Step Transformation | p. 112 |
| Cloning Vector for Desired Genes | p. 114 |
| Concluding Remarks | p. 115 |
| References | p. 116 |
| Safety Assessment of Insect Protected Crops: Testing the Feeding Value of Bt Corn and Cotton Varieties in Poultry, Swine and Cattle | |
| Introduction | p. 119 |
| Food Safety Standards | p. 119 |
| Testing for Food and Feed Safety | p. 120 |
| Insect Protection Traits | p. 122 |
| Benefits | p. 123 |
| Safety Assessment of the Cry Insect-Control Proteins | p. 124 |
| Mode of Action | p. 125 |
| Substantial Equivalence Based on Compositional Analysis | p. 126 |
| Current Products | p. 127 |
| Grower Acceptance | p. 127 |
| Future Products | p. 127 |
| Farm-Animal Studies | p. 128 |
| Bt Corn | p. 128 |
| Poultry | p. 128 |
| Lactating Cows | p. 131 |
| Beef and Sheep | p. 131 |
| Swine | p. 132 |
| Cottonseed | p. 133 |
| Conclusions | p. 134 |
| References | p. 135 |
| Safety Assessment of Genetically Modified Rice and Potatoes with Soybean Glycinin | |
| Introduction | p. 139 |
| Safety Assessment of Genetically Modified Crops | p. 140 |
| Genetically Modified Rice | p. 141 |
| Genetically Modified Potatoes | p. 144 |
| Concluding Remarks | p. 146 |
| References | p. 149 |
| Chromosomal and Genetic Aberrations in Transgenic Soybean | |
| Introduction | p. 153 |
| Times in Culture with 2,4-D Prior to Transformation | p. 154 |
| Genetic Background of the Explants | p. 159 |
| Seed Fertility in Transgenic Soybean | p. 160 |
| Cytological Basis of Gene Silencing | p. 163 |
| Conclusions | p. 164 |
| References | p. 165 |
| Transgenic Barley (Hordeum vulgare L.) and Chromosomal Variation | |
| Introduction | p. 169 |
| Chromosomal Variation in Nontransgenic Barley Plants | p. 170 |
| Chromosomal Variation in Transgenic Barley Plants | p. 172 |
| Fidelity and Quality of Transgenic Barley Plants | p. 177 |
| Comparative Analysis of Genomic Stability in Plants Derived from Tissues Generated Using Different in Vitro Proliferation Processes | p. 177 |
| Somaclonal Variation and Field Performance of Transgenic Plants Derived from Embryogenic Callus | p. 179 |
| Stability of Transgenes and Transgene Expression | p. 180 |
| Conclusions and Future Perspectives | p. 184 |
| References | p. 185 |
| Subject Index | p. 189 |
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