Practical Molecular Virology

ISBN-13: 9780896031913 / Angielski / Twarda / 1991 / 330 str.

Mary K. Collins

Opis

Recombinant viruses provide an efficient mechanism for the transfer and expression of DNA in eukaryotic cells. First, the transfer of DNA by viral infection-utilizing specific cell surface receptors and cellular intern- ization mechanisms-occurs much more readily than DNA transfer via uptake induced by such physical methods as calcium phosphate coprecipitation or electroporation. Second, the novel strategies employed by the virus to express its own genes can then be "hijacked" in the recombinant virus to express the researcher's gene of interest The purpose of Practical Molecular Virology isthus to compile a coll- tion of readily repeatable gene transfer and expression methods from wo- ers expert in the use of a variety of recombinant viral vectors . These include those designed for the production of recombinant antigens, such as pol- virus and yeast Ty-VLPs; those giving very high levels of recombinant protein expression, for example, baculovirus, vaccinia virus, and SV40; and finally viral vectors used for efficient, stable gene transfer to eu otic cells, such as retroviruses and herpesviruses . The first chapter describes the viral life cycle for each virus, and explains how this can be adapted to allow construction of recombinant vectors. Subsequent chapters deal with methods for producing and char- terizing recombinant viruses . I make no apology for the hyperproliferation of chapters dealing with recombinant retroviral methods and applications, since I believe this is clearly proportional to the recent expansion of interest in these techniques.

Recenzje

I found the writing style uniformly clear and straightforward throughout....I noticed the well-organized, parallel structure of most chapters, which made them easy to follow and gave evidence of some rather careful editing. Practical Molecular Virology provides a concise, well-organized, well-written...selective overview of eukaryotic viral vectors. As an entry-level text, it surely will assist those wishing to use such vectors to make a choice among established systems. Does this book meet its objective? I believe it does. Methods books are notoriously hard to execute well, but this one succeeds by carefully defining its own niche and then occupying it. -ASM News

Spis treści

The Retroviral Life Cycle and the Molecular Construction of Retrovirus Vectors. Construction of Retroviral Packaging Cell Lines. Making High Titer Retroviral Producer Cells. Methods of Titrating Nonselectable Recombinant Retroviruses. Selectable Markers for Eukaryotic Cells. Virus Genome Detection by the Polymerase Chain Reaction (PCR). Synctial Assays. Neutralization of Virus. Vesicular Stomatitis Virus Pseudotypes of Retroviruses. Cell Lineage Studies Using Retroviruses. Retroviral Vectors as Insertional Mutagents. Assessment of Compounds for Anti-HIV Activity. Analysis of Protein Factors that Interact with Adenovirus Early Promoters. COS Cell Expression. Papillomaviruses and Assays for Transforming Genes. Herpes Simplex Virus Life Cycle and the Design of Viral Vectors. Growth of Herpes Simplex Virus and Purification of Viral DNA. Vectors Based on Epstein-Barr Virus. Immortalization of Human B-Lymphocytes by Epstein-Barr Virus. Vaccinia Virus as an Expression Vector. Isolation and Handling of Recombinant Vaccinia Viruses. Design and Construction of Poliovirus Epitope Expression Vectors. Growth and Characterization of Poliovirus Antigen Chimeras. Expression Vectors for the Construction of Hybrid Ty-VLPs. Production and Purification of Hybrid Ty-VLPs. Baculovirus Expression Vectors: Choice of Expression Vector. Baculovirus Expression Vectors: Generating a Recombinant Baculovirus. Baculovirus Expression Vectors: Scale-Up and Downstream Processing. Index.