Genetic Engineering In Eukaryotes

This book includes the proceedings of a NATO Advanced Study Institute held at Washington State University, Pullman, Washington from July 26 until August 6, 1982. Although genetic engineering in eukaryotes is best developed in yeast and mammalian cells, the reader will find that some emphasis has been put on plant systems. Indeed, it was our position that the development of plant cell genetic transformation would benefit from the interactions between a comparatively smaller number of fungal and animal cell experts and a larger number of plant cell specialists representing various aspects of plant molecular genetic research. On the other hand, it is clear that the ultimate achievements of plant genetic engineering will have a tremendous ~pact on, among other things, food production without generating the problems of ethics encountered when one contempla-tes the genetic modification of human beings. Therefore, this slight bias in favor of the plant kingdom simply reflects our belief that a "second green revolution" will benefit mankind to a greater extent than any other kind of genetic engine ering. The keynote lecture of the Institute was delivered by Dr. John Slaughter, Director of the National Science Foundation, whom we deeply thank for his words of encouragement and commitment to the genetic manipulation of plants.

Discusses DNA including how it is put together, how cells read DNA, and the science and technology that is being explored based on cells and DNA.

Here's a concise introduction to all the key technologies of genetic engineering, including PCR, hybridization methods, gene mapping, cloning and targeting. A special section highlights the future of genetic engineering, including disease and gene therapies and the genetic engineering of animals and plants.

PART I Molecular Biology 1. Molecular Biology and Genetic Engineering Definition, History and Scope 2. Chemistry of the Cell: 1. Micromolecules (Sugars, Fatty Acids, Amino Acids, Nucleotides and Lipids) Sugars (Carbohydrates) 3. Chemistry of the Cell . 2. Macromolecules (Nucleic Acids; Proteins and Polysaccharides) Covalent and Weak Non-covalent Bonds 4. Chemistry of the Gene: Synthesis, Modification and Repair of DNA DNA Replication: General Features 5. Organisation of Genetic Material 1. Packaging of DNA as Nucleosomes in Eukaryotes Techniques Leading to Nucleosome Discovery 6. Organization of Genetic Material 2. Repetitive and Unique DNA Sequences 7. Organization of Genetic Material: 3. Split Genes, Overlapping Genes, Pseudogenes and Cryptic Genes Split Genes or .Interrupted Genes 8. Multigene Families in Eukaryotes 9. Organization of Mitochondrial and Chloroplast Genomes 10. The Genetic Code 11. Protein Synthesis Apparatus Ribosome, Transfer RNA and Aminoacyl-tRNA Synthetases Ribosome 12. Expression of Gene . Protein Synthesis 1. Transcription in Prokaryotes and Eukaryotes 13. Expression of Gene: Protein Synthesis: 2. RNA Processing (RNA Splicing, RNA Editing and Ribozymes) Polyadenylation of mRNA in Prokaryotes Addition of Cap (m7G) and Tail (Poly A) for mRNA in Eukaryotes 14. Expression of Gene: Protein Synthesis: 3. Synthesis and Transport of Proteins (Prokaryotes and Eukaryotes) Formation of Aminoacyl tRNA 15. Regulation of Gene Expression: 1. Operon Circuits in Bacteria and Other Prokaryotes 16. Regulation of Gene Expression . 2. Circuits for Lytic Cycle and Lysogeny in Bacteriophages 17. Regulation of Gene Expression 3. A Variety of Mechanisms in Eukaryotes (Including Cell Receptors and Cell Signalling) PART II Genetic Engineering 18. Recombinant DNA and Gene Cloning 1. Cloning and Expression Vectors 19. Recombinant DNA and Gene Cloning 2. Chimeric DNA, Molecular Probes and Gene Libraries 20. Polymerase Chain Reaction (PCR) and Gene Amplification 21. Isolation, Sequencing and Synthesis of Genes 22. Proteins: Separation, Purification and Identification 23. Immunotechnology 1. B-Cells, Antibodies, Interferons and Vaccines 24. Immunotechnology 2. T-Cell Receptors and MHC Restriction 25. Immunotechnology 3. Hybridoma and Monoclonal Antibodies (mAbs) Hybridoma Technology and the Production of Monoclonal Antibodies 26. Transfection Methods and Transgenic Animals 27. Animal and Human Genomics: Molecular Maps and Genome Sequences Molecular Markers 28. Biotechnology in Medicine: l.Vaccines, Diagnostics and Forensics Animal and Human Health Care 29. Biotechnology in Medicine 2. Gene Therapy Human Diseases Targeted for Gene Therapy Vectors and Other Delivery Systems for Gene Therapy 30. Biotechnology in Medicine: 3. Pharmacogenetics / Pharmacogenomics and Personalized Medicine Phannacogenetics and Personalized 31. Plant Cell and Tissue Culture' Production and Uses of Haploids 32. Gene Transfer Methods in Plants 33. Transgenic Plants . Genetically Modified (GM) Crops and Floricultural Plants 34. Plant Genomics: 35. Genetically Engineered Microbes (GEMs) and Microbial Genomics References

Discusses current and potential uses of genetic engineering in fields such as medicine, criminal investigation, and agriculture and examines some of the ethical questions involved.

Molecular Genetic Modification of Eucaryotes reviews the current state of knowledge and techniques potentially useful for the molecular genetic modification of eucaryotes. The book interfaces molecular genetics, plant cell and tissue culture, and plant improvement. Topics include cell and tissue culture, protoplasts, somatic cell fusion, cellular mutagenesis, regeneration of new plant types, and the applicability of these techniques to plant improvements. Viruses and viral integration; integration and expression of foreign genetic material in human cells, Drosophila, plant cells, and legumes; biophysical studies of DNA uptake by plants; and genetic engineering for plant protection against diseases are also discussed in detail. The book will be of interest to geneticists, cell biologists, plant breeders, plant physiologists, plant pathologists, and biochemists.

The possession of plasmids was for a long time recognized only in the bacteria. It is now evident that plasmids, or replicative forms of DNA structurally and experimentally comparable to bacterial plasmids, exist in eukaryotic organisms as well. Such plasmids are in fact common among fungi and higher plants. The present review is undertaken to provide a comprehensive account of the data available on plasmids found in eukaryotic organisms. This review will not consider plasmids of prokaryotic origin, even though certain bacterial plasmids, such as the tumor-inducing (Ti) plasmids of Agrobacterium tumefaciens, may be intimately associated with transformation of the eukaryotic host. This book, moreover, does not consider transformation experiments in eukaryotic hosts involving viral DNA as vectors, although indeed such vectors have been developed for use in plant and animal systems. After a general introduction, providing historical perspective on the nature and role of plasmids, a list of eukaryotic plasmids will be presented according to their origin. This is followed by a detailed discussion of known structure and function. In subsequent chapters the practical implications of eukaryotic plasmids for molecular cloning and biotechnology will be discussed. This latter part traces the development of interest'in biotechnical genetics and gives special consideration to the use of eukaryotic systems for gene cloning. The terminology biotechni cal genetics is introduced to the reader and is used in a general sense as equivalent to genetic engineering. Biotechnical genetics includes, but is not limited to, gene cloning through recombinant DNA technology.