A major success story of modem molecular biology is the development of technologies to clone and express specific genes. Current applications of recombinant gene products cover a wide spectrum, including gene therapy, production of bioactive pharmaceuticals, synthesis of novel biopolymers, agriculture and animal husbandry, and so on. Inherent in bringing these appli cations to fruition is the need to design "expression constructs" that will per mit the ready and specific detection and isolation of the defined recombinant gene products. Recombinant Protein Protocols grows out of the need for a laboratory manual on the detection and isolation of recombinantly expressed genes that covers both the background information and the practical laboratory recipes for these analyses. In this book, detailed and contemporary protocols are col lected to provide the reader with a wide-ranging number of methodologies to enhance the detection and isolation of their gene product(s) of interest. A large number of molecular tags and labels and their usage are described, including enzymes, ligand-binding moieties, immunodetectable molecules, as well as methods to detect interactive proteins, and gene expression-mediated alter ations in cellular activity. Chapters on in situ detection of gene expression deal with technologies that are currently being applied to the study of gene function and activity. Highlights of applications for recombinant gene expres sion technologies are provided to give readers exciting perspectives on the future of such technologies.

The current explosive progress in molecular biological research can be definitively traced to the development of molecular cloning technology. The ability to insert specific gene sequences into cloning vectors and their subse quent expansion is the cornerstone of modem molecular biology. A direct practical outcome of molecular cloning technology is its application to ex press specific recombinant genes. Currently, recombinant gene products are used in a wide spectrum of applications, including gene therapy, production of bioactive pharmaceuticals, synthesis of novel biopolymers, in agriculture and animal husbandry, and so on. A fundamental requirement for successful recombinant gene expression is the design of the cloning vector and the choice of the host organism for expression. Recombinant Gene Expression Protocols grows out of the need for a laboratory manual that provides the reader the background and rationale, as well as the practical protocols for the preparation of "expression constructs" and their introduction into appropriate host cells and/or organisms. The chap ters in this book are grouped by their expression hosts, including E. coli, yeast, mammalian cells, nonmammalian eukaryotes such as plants, Xenopus, and insects, as well as in transgenic organisms. In-depth information is presented on the important characteristics of expression cloning vectors and the various methods for efficiently introducing expression constructs into target cells and/ or organisms. Throughout Recombinant Gene Expression Protocols, the authors have consistently striven for a balanced presentation of both background informa tion and actual laboratory details.

Drawing on state-of-the-art cellular and molecular techniques as well as new and sophisticated imaging and information technologies, this comprehensive, three-volume collection of cutting-edge protocols provides readily reproducible methods for studying and analyzing the events of embryonic development. Volume 1 (ISBN: 089603-574-3) contains techniques for establishing and characterizing several widely used experimental model systems, for the study of developmental patterns and morphogenesis, and for the examination of embryo structure and function. There are also step-by-step methods for the analaysis of cell lineage, the production and use of chimeras, and the experimental and molecular manipulation of embryos, including the application of viral vectors. Volume 2 (ISBN: 0-89603-575-1) describes state-of-the-art methods for the study of organogenesis, the analysis of abnormal development and teratology, the screening and mapping of novel genes and mutations, and the application of transgenesis, including the production of transgenic animals and gene knockouts. No less innovative, Volume 3 (ISBN: 0-89603-576-X) introduces powerful techniques for the manipulation of developmental gene expression and function, the analysis of gene expression, the characterization of tissue morphogenesis and development, the in vitro study of differentiation and development, and the genetic analysis of developmental models of diseases. Highly practical and richly annotated, the three volumes of Developmental Biology Protocols describe multiple experimental systems and details techniques adopted from the broadest array of biomedical disciplines.

Drawing on state-of-the-art cellular and molecular techniques as well as new and sophisticated imaging and information technologies, this comprehensive, three-volume collection of cutting-edge protocols provides readily reproducible methods for studying and analyzing the events of embryonic development. Volume 1 (ISBN: 089603-574-3) contains techniques for establishing and characterizing several widely used experimental model systems, for the study of developmental patterns and morphogenesis, and for the examination of embryo structure and function. There are also step-by-step methods for the analaysis of cell lineage, the production and use of chimeras, and the experimental and molecular manipulation of embryos, including the application of viral vectors. Volume 2 (ISBN: 0-89603-575-1) describes state-of-the-art methods for the study of organogenesis, the analysis of abnormal development and teratology, the screening and mapping of novel genes and mutations, and the application of transgenesis, including the production of transgenic animals and gene knockouts. No less innovative, volume 3 (ISBN: 0-89603-576-X) introduces powerful techniques for the manipulation of developmental gene expression and function, the analysis of gene expression, the characterization of tissue morphogenesis and development, the in vitro study of differentiation and development, and the genetic analysis of developmental models of diseases. Highly practical and richly annotated, the three volumes of Developmental Biology Protocols describe multiple experimental systems and details techniques adopted from the broadest array of biomedical disciplines.

Drawing on state-of-the-art cellular and molecular techniques as well as new and sophisticated imaging and information technologies, this comprehensive, three-volume collection of cutting-edge protocols provides readily reproducible methods for studying and analyzing the events of embryonic development. Volume 1 (ISBN: 089603-574-3) contains techniques for establishing and characterizing several widely used experimental model systems, for the study of developmental patterns and morphogenesis, and for the examination of embryo structure and function. There are also step-by-step methods for the analaysis of cell lineage, the production and use of chimeras, and the experimental and molecular manipulation of embryos, including the application of viral vectors. Volume 2 (ISBN: 0-89603-575-1) describes state-of-the-art methods for the study of organogenesis, the analysis of abnormal development and teratology, the screening and mapping of novel genes and mutations, and the application of transgenesis, including the production of transgenic animals and gene knockouts. No less innovative, Volume 3 (ISBN: 0-89603-576-X) introduces powerful techniques for the manipulation of developmental gene expression and function, the analysis of gene expression, the characterization of tissue morphogenesis and development, the in vitro study of differentiation and development, and the genetic analysis of developmental models of diseases. Highly practical and richly annotated, the three volumes of Developmental Biology Protocols describe multiple experimental systems and details techniques adopted from the broadest array of biomedical disciplines.

A major success story of modem molecular biology is the development of technologies to clone and express specific genes. Current applications of recombinant gene products cover a wide spectrum, including gene therapy, production of bioactive pharmaceuticals, synthesis of novel biopolymers, agriculture and animal husbandry, and so on. Inherent in bringing these appli cations to fruition is the need to design "expression constructs" that will per mit the ready and specific detection and isolation of the defined recombinant gene products. Recombinant Protein Protocols grows out of the need for a laboratory manual on the detection and isolation of recombinantly expressed genes that covers both the background information and the practical laboratory recipes for these analyses. In this book, detailed and contemporary protocols are col lected to provide the reader with a wide-ranging number of methodologies to enhance the detection and isolation of their gene product(s) of interest. A large number of molecular tags and labels and their usage are described, including enzymes, ligand-binding moieties, immunodetectable molecules, as well as methods to detect interactive proteins, and gene expression-mediated alter ations in cellular activity. Chapters on in situ detection of gene expression deal with technologies that are currently being applied to the study of gene function and activity. Highlights of applications for recombinant gene expres sion technologies are provided to give readers exciting perspectives on the future of such technologies.

Developmental biology is one of the most exciting and fast-growing fields today. In part, this is so because the subject matter deals with the innately fascinating biological events-changes in form, structure, and function of the organism. The other reason for much of the excitement in developmental biology is that the field has truly become the unifying melting pot of biology, and provides a framework that integrates anatomy, physiology, genetics, biochemistry, and cellular and molecular biology, as well as evolutionary biology. No longer is the study of embryonic development merely "embryology." In fact, development biology has produced important paradigms for both basic and clinical biomedical sciences. Though modern developmental biology has its roots in "experimental embry- ogy" and the even more classical "chemical embryology," the recent explosive and remarkable advances in developmental biology are critically linked to the advent of the "cellular and molecular biology revolution." The impressive arsenal of expe- mental and analytical tools derived from cell and molecular biology, which promise to continue to expand, together with the exponentially developing sophistication in fu- tional imaging and information technologies, guarantee that the study of the devel- ing embryo will contribute one of the most captivating areas of biological research in the next millennium.

Drawing on state-of-the-art cellular and molecular techniques as well as new and sophisticated imaging and information technologies, this comprehensive, three-volume collection of cutting-edge protocols provides readily reproducible methods for studying and analyzing the events of embryonic development. volume 1 (ISBN: 089603-574-3) contains techniques for establishing and characterizing several widely used experimental model systems, for the study of developmental patterns and morphogenesis, and for the examination of embryo structure and function. There are also step-by-step methods for the analaysis of cell lineage, the production and use of chimeras, and the experimental and molecular manipulation of embryos, including the application of viral vectors. volume 2 (ISBN: 0-89603-575-1) describes state-of-the-art methods for the study of organogenesis, the analysis of abnormal development and teratology, the screening and mapping of novel genes and mutations, and the application of transgenesis, including the production of transgenic animals and gene knockouts. No less innovative, volume 3 (ISBN: 0-89603-576-X) introduces powerful techniques for the manipulation of developmental gene expression and function, the analysis of gene expression, the characterization of tissue morphogenesis and development, the in vitro study of differentiation and development, and the genetic analysis of developmental models of diseases. Highly practical and richly annotated, the three volumes of Developmental Biology Protocols describe multiple experimental systems and details techniques adopted from the broadest array of biomedical disciplines.

Drawing on state-of-the-art cellular and molecular techniques as well as new and sophisticated imaging and information technologies, this comprehensive, three-volume collection of cutting-edge protocols provides readily reproducible methods for studying and analyzing the events of embryonic development. volume 1 (ISBN: 089603-574-3) contains techniques for establishing and characterizing several widely used experimental model systems, for the study of developmental patterns and morphogenesis, and for the examination of embryo structure and function. There are also step-by-step methods for the analaysis of cell lineage, the production and use of chimeras, and the experimental and molecular manipulation of embryos, including the application of viral vectors. volume 2 (ISBN: 0-89603-575-1) describes state-of-the-art methods for the study of organogenesis, the analysis of abnormal development and teratology, the screening and mapping of novel genes and mutations, and the application of transgenesis, including the production of transgenic animals and gene knockouts. No less innovative, volume 3 (ISBN: 0-89603-576-X) introduces powerful techniques for the manipulation of developmental gene expression and function, the analysis of gene expression, the characterization of tissue morphogenesis and development, the in vitro study of differentiation and development, and the genetic analysis of developmental models of diseases. Highly practical and richly annotated, the three volumes of Developmental Biology Protocols describe multiple experimental systems and details techniques adopted from the broadest array of biomedical disciplines.

Drawing on state-of-the-art cellular and molecular techniques as well as new and sophisticated imaging and information technologies, this comprehensive, three-volume collection of cutting-edge protocols provides readily reproducible methods for studying and analyzing the events of embryonic development. Volume 1 (ISBN: 089603-574-3) contains techniques for establishing and characterizing several widely used experimental model systems, for the study of developmental patterns and morphogenesis, and for the examination of embryo structure and function. There are also step-by-step methods for the analaysis of cell lineage, the production and use of chimeras, and the experimental and molecular manipulation of embryos, including the application of viral vectors. Volume 2 (ISBN: 0-89603-575-1) describes state-of-the-art methods for the study of organogenesis, the analysis of abnormal development and teratology, the screening and mapping of novel genes and mutations, and the application of transgenesis, including the production of transgenic animals and gene knockouts. No less innovative, volume 3 (ISBN: 0-89603-576-X) introduces powerful techniques for the manipulation of developmental gene expression and function, the analysis of gene expression, the characterization of tissue morphogenesis and development, the in vitro study of differentiation and development, and the genetic analysis of developmental models of diseases. Highly practical and richly annotated, the three volumes of Developmental Biology Protocols describe multiple experimental systems and details techniques adopted from the broadest array of biomedical disciplines.

The current explosive progress in molecular biological research can be definitively traced to the development of molecular cloning technology. The ability to insert specific gene sequences into cloning vectors and their subse quent expansion is the cornerstone of modem molecular biology. A direct practical outcome of molecular cloning technology is its application to ex press specific recombinant genes. Currently, recombinant gene products are used in a wide spectrum of applications, including gene therapy, production of bioactive pharmaceuticals, synthesis of novel biopolymers, in agriculture and animal husbandry, and so on. A fundamental requirement for successful recombinant gene expression is the design of the cloning vector and the choice of the host organism for expression. Recombinant Gene Expression Protocols grows out of the need for a laboratory manual that provides the reader the background and rationale, as well as the practical protocols for the preparation of "expression constructs" and their introduction into appropriate host cells and/or organisms. The chap ters in this book are grouped by their expression hosts, including E. coli, yeast, mammalian cells, nonmammalian eukaryotes such as plants, Xenopus, and insects, as well as in transgenic organisms. In-depth information is presented on the important characteristics of expression cloning vectors and the various methods for efficiently introducing expression constructs into target cells and/ or organisms. Throughout Recombinant Gene Expression Protocols, the authors have consistently striven for a balanced presentation of both background informa tion and actual laboratory details.