Gene Targeting

A Practical Approach
2. Auflage 1999
ISBN: 978-0-19-963792-8
Verlag: Oxford University Press

Buch, Englisch, 312 Seiten, Print PDF, Format (B × H): 158 mm x 234 mm, Gewicht: 483 g

Since the publication of the first edition of Gene Targeting: A Practical Approach in 1993 there have been many advances in gene targeting and this new edition has been thoroughly updated and rewritten to include all the major new techniques. It provides not only tried-and-tested practical protocols but detailed guidance on their use and applications. As with the previous edition Gene Targeting: A Practical Approach 2e concentrates on gene targeting in mouse ES cells, but the techniques described can be easily adapted to applications in tissue culture including those for human cells. The first chapter covers the design of gene targeting vectors for mammalian cells and describes how to distinguish random integrations from homologous recombination. It is followed by a chapter on extending conventional gene targeting manipulations by using site-specific recombination using the Cre-loxP and Flp-FRT systems to produce 'clean' germline mutations and conditionally (in)activating genes. Chapter 3 describes methods for introducing DNA into ES cells for homologous recombination, selection and screening procedures for identifying and recovering targeted cell clones, and a simple method for establishing new ES cell lines. Chapter 4 discusses the pros and cons or aggregation versus blastocyst injection to create chimeras, focusing on the technical aspects of generating aggregation chimeras and then describes some of the uses of chimeras. The next topic covered is gene trap strategies; the structure, components, design, and modification of GT vectors, the various types of GT screens, and the molecular analysis of GT integrations. The final chapter explains the use of classical genetics in gene targeting and phenotype interpretation to create mutations and elucidate gene functions. Gene Targeting: A Practical Approach 2e will therefore be of great value to all researchers studying gene function.

Autoren/Hrsg.

Joyner, Alexandra

Fachgebiete

• Naturwissenschaften Biowissenschaften Proteinforschung
• Technische Wissenschaften Verfahrenstechnik | Chemieingenieurwesen | Biotechnologie Biotechnologie Medizinische Biotechnologie
• Naturwissenschaften Biowissenschaften Biowissenschaften DNA und Transgene Organismen
• Naturwissenschaften Biowissenschaften Molekularbiologie

Inhaltsverzeichnis

- Preface

- 1: Bradley et al: Gene targeting vectors for mammalian cells

- 2: Dymecki: Site specific recombination in cell and mice

- 3: Matise and Joyner: Production of targeted ES cell clones

- 4: Papaioannou and Johnson: Production of chimeras and genetically defined offspring from targeted ES cells

- 5: Nagy and Rossant: Production of completely ES cell-derived fetuses

- 6: Wurst and Gossler: Gene and enhancer traps in ES cells

- 7: Bultman and Magnuson: Mouse genetics and analysis of mutant phenotypes

- Index