Transgenesis Techniques

by Clarke, Alan R.

• ISBN13: 9780896036963
• ISBN10: 0896036960
• Edition: 2nd
• Format: Hardcover
• Copyright: 2002-01-01
• Publisher: Humana Pr Inc

Summary

This revised and updated edition of a much-acclaimed classic presents a collection of readily reproducible techniques for the generation and analysis of transgenic animals, with emphasis on the manipulation of the mammalian genome. It concentrates on the creation and maintenance of genetically modified murine strains, providing access to the germline by both conventional pronuclear injection, and by retroviral and adenoviral infection. Extensive coverage is also given to the generation, maintenance, and manipulation of embryonic stem cell lineages, with protocols for both constitutive and conditional (Cre-lox or the Flp-frt systems) gene targeting. Additional chapters provide cutting-edge techniques for the cryopreservation of both male and female germlines, for the generation of transgenic sheep by nuclear transfer, and for the basic analysis of transgenic organisms.

Table of Contents

Preface	v
Contributors	ix
Part I. Topical Reviews in Transgenesis
Biomedical and Agricultural Applications of Animal Transgenesis	3	(24)
Jim McWhir
Part II. Transgenesis in Invertebrate and Lower Vertebrate Species
Gene Transfer in Drosophila	27	(12)
Mark J. O'Connor
William Chia
Part III. Transgenesis in the Mouse: Oocyte Injection
Oocyte Injection in the Mouse	39	(34)
Gary A. J. Brown
Timothy J. Corbin
Part IV. Alternative Routes to the Germline
Adenoviral Infection	73	(10)
Tohru Tsukui
Yutaka Toyodo
Retroviral Infection	83	(10)
Richard A. Bowen
Part V. Transgenesis in the Mouse: the Es Cell Route
In Vitro Isolation of Murine Embryonic Stem Cells	93	(34)
David Wells
Production of Chimeras Derived from Murine Embryonic Stem Cells	127	(24)
David Wells
Gene Targeting Strategies	151	(24)
David W. Melton
Cre/IoxP Recombination System and Gene Targeting	175	(32)
Ralf Kuhn
Raul M. Torres
Part VI. Cryopreservation of Mouse Lines
Cryopreservation of Transgenic Mouse Lines	207	(22)
Jillian M. Shaw
Naomi Nakagata
Ovarian Tissue Transplantation and Cryopreservation: Application to Maintenance and Recovery of Transgenic and Inbred Mouse Lines	229	(26)
Jillian M. Shaw
A. O. Trounson
Part VII. Transgenesis in the Rat
Transgenesis in the Rat	255	(18)
Linda J. Mullins
Gillian Brooker
John J. Mullins
Part VIII. Transgenesis in Domestic Species
Generation of Transgenic Livestock by Pronuclear Injection	273	(16)
A. John Clark
Transgenic Sheep from Cultured Cells	289	(16)
Keith H. S. Campbell
Part IX. Characterization and Analysis of Transgenic Strains
Analysis of Transgenic Mice	305	(38)
Stefan Selbert
Dominic Rannie
Index	343