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Transgenesis Techniques Principles and Protocols / by David Murphy, David A. Carter.

Por: Murphy, David, autor.
Colaborador(es): Carter, David A, autor.
Series (Methods in Molecular Biology, 1940-6029; 18).Editor: Totowa, NJ : Humana Press, 1993Edición: 1st edition 1993.Descripción: 1 recurso en línea (XII, 467 páginas) : .ISBN: 9781592595051.Recursos en línea: (usuarios Universidad Europea de Valencia)Digital Resources
Contenidos:
and Topical Reviews -- to Transgenesis -- Transgenic Rodents and the Study of the Central Nervous System -- Transgenic Animals and the Study of Cancer -- Transgenic Animals and the Study of the Immune System -- The Commercial and Agricultural Applications of Animal Transgenesis -- Transgenesis in Invertebrate and Lower Vertebrate Species -- P Element-Mediated Germ-Line Transformation of Drosophila -- Gene Transfer by Microinjection in the Zebra Fish Brachydanio rerio -- Transgenic Induction in Salmonid and Tilapia Fish -- Transgenesis in the Mouse -- An Overview of Transgenic Mouse Production -- Transgene Design -- Isolation of DNA Fragments for Microinjection -- Establishing a Colony for Efficient Production of Transgenic Mice -- Mating Mice -- Anesthetizing Mice -- Vasectomizing a Mouse -- Preparation of Culture Media for Fertilized One-Cell Mouse Eggs -- Collection of Fertilized One-Cell Mouse Eggs for Microinjection -- Microinjection of Cloned DNA Fragments into Fertilized One-Cell Mouse Eggs -- Microinjection of Cloned DNA Fragments into Fertilized One-Cell Mouse Eggs -- Delivery of Microinjected Eggs to Surrogate Mothers by Oviduct Transfer -- Caesarean Section and Fostering -- In vitro Fertilization of Mouse Eggs -- The In Vitro Isolation of Murine Embryonic Stem Cells -- Production of Chimeras Derived from Murine Embryonic Stem Cells -- Cryopreservation of Transgenic Mouse Lines -- Transgenesis in the Rat -- to the Physiology and Husbandry of the Rat -- Mating of Rats -- Vasectomy of the Male Rat -- Harvest of Fertilized One-Cell Rat Eggs -- Implantation of Microinjected Eggs -- Cryopreservation of Transgenic Rat Lines -- Transgenesis in a Domestic Species-The Sheep -- Production of Transgenic Sheep -- Cryopreservation of Transgenic Sheep Lines -- Genomic Analysis of Transgenic Animals -- Isolation of Genomic DNA from Tail Tissue -- Slot Blotting of Genomic Tail DNA -- Genomic Analysis of Transgenic Animals by the Polymerase Chain Reaction -- Genomic Analysis of Transgenic Animals -- Expression Analysis of Transgenic Animals -- The Preparation of RNA from the Tissues of Transgenic Animals -- Northern Blotting -- Rna slot Blotting -- Analysis of Gene Expression by PCR -- Nuclear Run-On Analysis of Transcription -- S1 Nuclease Protection Mapping -- Primer Extension -- In Situ Hybridization Analysis of Transgenic Tissues -- Immunohistochemical Analysis of Transgene Expression -- Postmortem Examination of Transgenic Mice -- Histological Examination of Transgenic Mice -- Reporter Enzyme Assays -- Restriction Endonuclease Digestion of DNA -- Agarose Gel Electrophoresis -- Working with Radioactivity in the Transgenic Animal Laboratory -- Random Primed Labeling of DNA -- Filter Hybridization -- DNA Transfection.
Resumen: Over the past twenty years, the disciplines of cellular and molecular biology have generated a bounty of information pertaining to the understanding of the structure and function of eukaryotic cells . It is now recognized that individual cells regulate their activities in response to a multitude of external signals ; that external signals, a- ing through a limited number of second messenger systems, modulate the cell's pattern of gene expression; and that the cohort of genes available for regulation is dependent upon the developmental history of the cell. Extrapolating these truths to the whole organism is a challenge that has been met by an increasing number of investigators . The wealth of information that molecular and cell biology has provided us with is now being integrated into an understanding of whole t- sues, organs, and organisms. Without such an integration, the molecular information will be nothing more than a confusing catalog of sequences and structures. Central to this project will be the app- cation of the powerful new techniques described in Transgenesis Techniques: Principles and Protocols .
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and Topical Reviews -- to Transgenesis -- Transgenic Rodents and the Study of the Central Nervous System -- Transgenic Animals and the Study of Cancer -- Transgenic Animals and the Study of the Immune System -- The Commercial and Agricultural Applications of Animal Transgenesis -- Transgenesis in Invertebrate and Lower Vertebrate Species -- P Element-Mediated Germ-Line Transformation of Drosophila -- Gene Transfer by Microinjection in the Zebra Fish Brachydanio rerio -- Transgenic Induction in Salmonid and Tilapia Fish -- Transgenesis in the Mouse -- An Overview of Transgenic Mouse Production -- Transgene Design -- Isolation of DNA Fragments for Microinjection -- Establishing a Colony for Efficient Production of Transgenic Mice -- Mating Mice -- Anesthetizing Mice -- Vasectomizing a Mouse -- Preparation of Culture Media for Fertilized One-Cell Mouse Eggs -- Collection of Fertilized One-Cell Mouse Eggs for Microinjection -- Microinjection of Cloned DNA Fragments into Fertilized One-Cell Mouse Eggs -- Microinjection of Cloned DNA Fragments into Fertilized One-Cell Mouse Eggs -- Delivery of Microinjected Eggs to Surrogate Mothers by Oviduct Transfer -- Caesarean Section and Fostering -- In vitro Fertilization of Mouse Eggs -- The In Vitro Isolation of Murine Embryonic Stem Cells -- Production of Chimeras Derived from Murine Embryonic Stem Cells -- Cryopreservation of Transgenic Mouse Lines -- Transgenesis in the Rat -- to the Physiology and Husbandry of the Rat -- Mating of Rats -- Vasectomy of the Male Rat -- Harvest of Fertilized One-Cell Rat Eggs -- Implantation of Microinjected Eggs -- Cryopreservation of Transgenic Rat Lines -- Transgenesis in a Domestic Species-The Sheep -- Production of Transgenic Sheep -- Cryopreservation of Transgenic Sheep Lines -- Genomic Analysis of Transgenic Animals -- Isolation of Genomic DNA from Tail Tissue -- Slot Blotting of Genomic Tail DNA -- Genomic Analysis of Transgenic Animals by the Polymerase Chain Reaction -- Genomic Analysis of Transgenic Animals -- Expression Analysis of Transgenic Animals -- The Preparation of RNA from the Tissues of Transgenic Animals -- Northern Blotting -- Rna slot Blotting -- Analysis of Gene Expression by PCR -- Nuclear Run-On Analysis of Transcription -- S1 Nuclease Protection Mapping -- Primer Extension -- In Situ Hybridization Analysis of Transgenic Tissues -- Immunohistochemical Analysis of Transgene Expression -- Postmortem Examination of Transgenic Mice -- Histological Examination of Transgenic Mice -- Reporter Enzyme Assays -- Restriction Endonuclease Digestion of DNA -- Agarose Gel Electrophoresis -- Working with Radioactivity in the Transgenic Animal Laboratory -- Random Primed Labeling of DNA -- Filter Hybridization -- DNA Transfection.

Over the past twenty years, the disciplines of cellular and molecular biology have generated a bounty of information pertaining to the understanding of the structure and function of eukaryotic cells . It is now recognized that individual cells regulate their activities in response to a multitude of external signals ; that external signals, a- ing through a limited number of second messenger systems, modulate the cell's pattern of gene expression; and that the cohort of genes available for regulation is dependent upon the developmental history of the cell. Extrapolating these truths to the whole organism is a challenge that has been met by an increasing number of investigators . The wealth of information that molecular and cell biology has provided us with is now being integrated into an understanding of whole t- sues, organs, and organisms. Without such an integration, the molecular information will be nothing more than a confusing catalog of sequences and structures. Central to this project will be the app- cation of the powerful new techniques described in Transgenesis Techniques: Principles and Protocols .

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