Description |
xx, 521 pages : illustrations (some color) ; 24 cm |
Series |
Methods in molecular biology ; v. 62 |
|
Methods in molecular biology (Clifton, N.J.) ; v. 62
|
Contents |
1. Introduction -- Recombinanat gene expression protocols overview of experimental strategies for the expression of recombinant genes -- 2. Expression in E. coli -- Designing expression plasmid vectors in E. coli -- Design of bacterial hosts for Iac-based expressed vectors -- Bacteriophage gamma-based expression vectors -- Strategies for gene fusions -- Application of the E. coli trp promoter -- Expression of proteins in E. coli utilizing a dual promoter-based vector: pLACT7 -- Expression and secretion of proteins in E. coli -- Synthetic two-cistron expression system -- Expression of cDNA-encoded proteins by cell-free transcription and translation -- 3. Expression in yeast -- Yeast plasids -- Inducible expression cassettes in yeast: GAL4 -- Inducible expression cassettes in yeast: ADH2 -- Constitutive expression vectors: PGK -- 4. Viral expression systems -- Design and construction of recombinant vaccinia viruses -- A simplified method of screening for isolation of recombinant vaccinia virus -- Design of retroviral expression vectors -- Use of defective herpes-derived plasmid vectors -- 5. Expression in nonmammalian higher eukaryotes -- Use of baculovirus expression vectors -- Expression of exogenous genes in Xenopus oocytes, eggs, and embryos -- Analysis of heterologous gene expression in Xenopus blastomeres -- 6. Expression in mammalian cells -- Overview of vector design for mammalian gene expression -- Experimental strategies in efficient transfection of mammalian cells: calcium phosphate and DAAE-dextran -- Experimental strategies in efficient transfection of mammalian cells: electroporation -- Highly effective delivery of foreign DNA to atherent cells via polybrene/DMSO-assisted gene transfer -- Selection of transfected cells: magnetic affinity cell sorting -- Selection of transfected cells and coamplification of transfected genes -- Optimization of growth, viability, and specific productivity for expression of recombinant proteins in mammalian cells -- 7. Expression in transgenic organisms -- 8. Expression in Plants |
Analysis |
Genetic recombination Laboratory manuals |
|
Gene Expression |
|
Genetic Engineering |
|
Laboratory Manuals |
|
Recombination, Genetic |
Notes |
Includes index |
Bibliography |
Includes bibliographical references and index |
Notes |
Also available online via the World Wide Web, by subscription to BioMedProtocols.com (BMP) via Humana Press |
|
Methods in molecular biology (Clifton, N.J.) no:62 1064-3745 |
Subject |
Genetic recombination -- Laboratory manuals.
|
|
Recombination, Genetic.
|
|
Gene Expression.
|
|
Genetic Engineering.
|
Genre/Form |
Laboratory Manual.
|
|
Laboratory manuals.
|
Author |
Tuan, Rocky S.
|
|
Humana Press.
|
LC no. |
96049600 |
ISBN |
0896033333 (alk. paper) |
|