Microinjection and Transgenesis
Springer Berlin (Verlag)
978-3-540-61895-9 (ISBN)
The establishment of microinjection protocols about 20 years ago for cultured cells and shortly thereafter for the generation of transgenic mice by microinjection of DNA into fertilized mouse eggs greatly influ enced many fields of biology. Not only have the data generated using these approaches contributed to a large extent to our present under standing of gene regulation and cellular function of higher eukaryotic cells, but current knowledge and future developments in this area will certainly have a great impact on basic and applied research for many years to come. This laboratory manual describes the current state of the art in this research area and focuses primarily on both the experimental strategies with an extensive bibliography and the detailed procedures. A large number of studies are presently being performed and a great variety of new experimental designs are rapidly being developed. The book con tains protocols on injection of somatic cells as well as on injection of embryos, the use of similar equipment being a common feature. In the articles dedicated to somatic cells, full descriptions of the manual and automatic injection systems are given as well as the methods for the analysis of injected cells by video-microscopy, electron microscopy or in situ hybridizations. In addition, comprehensive protocols are given for injection experiments with very different purposes, such as to study sig nal transduction or microtubule dynamics.
1 Strategies and Protocols for Microinjection Experiments Using Somatic Cells in Culture.- 1 Protocols for the Manual and Automatic Microinjection of Somatic Cells in Culture and for the Analysis of Microinjected Cells.- 2 Automated Computer-Assisted Microinjection into Cultured Somatic Cells.- 3 Immunofluorescence and Immunoelectron Microscopy of Microinjected and Transfected Cultured Cells.- 4 Videomicroscopy of Living Microinjected Hippocampal Neurons in Culture.- 5 Preparation, Purification, and Analysis of Synthetic Oligonucleotides Suitable for Microinjection Experiments.- 6 Colorimetric in Situ Hybridization.- 7 Manual Microinjection: Measuring the Inhibitory Effect of HIV-1-Directed Hammerhead Ribozymes in Tissue Culture Cells.- 8 Cellular Microbiochemistry.- 9 A New Strategy for Studying Microtubule Transport and Assembly During Axon Growth.- 10 The Use of Microinjection to Study Signal Transduction in Mammalian Cells.- 2 Strategies and Protocols for Microinjection Experiments Using Embryos.- 11 Supply and Husbandry of Mice for Transgenic Science.- 12 Mouse Male Germ Cells in Culture: Toward a New Approach in Transgenesis?.- 13 Sperm-Mediated Gene Transfer.- 14 Application of Ligand-Dependent Site-Specific Recombination in ES Cells.- 15 A Modified Protocol for the Generation of Transgenic Mice and Freezing of Mouse Embryos.- 16 The Use of Morula Aggregation to Generate Germline Chimeras from Genetically Modified Embryonic Stem Cells.- 17 The Generation of Transgenic Mice with Yeast Artificial Chromosomes.- 18 Following Up Microinjected mRNAs: Analysis of mRNA Poly(A) Tail Length During Mouse Oocyte Maturation.- 19 Mouse Preimplantation Embryos and Oocytes as an In Vivo System to Study Transcriptional Enhancers.- 20 The Use of Mouse Preimplantation Embryos for the Identification of DNA-Binding Proteins Essential in Early Development.- 21 Application of Transgenes and Transgenic Mice to Study Gene Activity from the Oocyte to the Early Embryo.- 22 Transitory Transgenic Analysis as an In Vivo System to Study Promoter Regulatory Elements.- 23 Transgenic Analysis of Embryonic Gene Expression Using LacZ as a Reporter.- 24 Autonomous Cell Labeling Using a laacZ Reporter Transgene to Produce Genetic Mosaics During Development.- 25 Multiple Developmental Functions of the Zinc Finger Gene Krox-20 Are Revealed by Targeted Insertion of the E. coli lacZ Coding Sequence.- 26 Transgenic Mouse Strategies in Virus Research.- 27 Isolation of Mutated Genes from Transgene Insertion Sites.- 28 Contributions of Transgenic and Knockout Mice to Immunological Knowledge.- 29 Generation and Application of Transgenic Rabbits.- 30 Microinjection of Drosophila Eggs.
| Erscheint lt. Verlag | 27.11.1997 |
|---|---|
| Reihe/Serie | Springer Lab Manuals |
| Zusatzinfo | XIV, 619 p. 66 illus., 10 illus. in color. |
| Verlagsort | Berlin |
| Sprache | englisch |
| Maße | 193 x 242 mm |
| Gewicht | 1268 g |
| Themenwelt | Naturwissenschaften ► Biologie ► Mikrobiologie / Immunologie |
| Naturwissenschaften ► Biologie ► Zellbiologie | |
| Schlagworte | biochemistry • Cell • Cell Culture • chromosome • electron microscopy • Gene • gene expression • gene transfer • Genmanipulation • Klonieren • Microscopy • Mikroinjektion • Mutation • proteins • Ribozyme • Stem Cells • tissue • transcription • Transgene • Transgene Mäuse • Transgenice mice • Vivo • Yeast artificial chromosome • Zellinjektion • Zellkultur |
| ISBN-10 | 3-540-61895-3 / 3540618953 |
| ISBN-13 | 978-3-540-61895-9 / 9783540618959 |
| Zustand | Neuware |
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