Sv40 Protocols

ISBN-13: 9781489942074 / Angielski / Miękka / 2013 / 310 str.

Leda Raptis

Opis

Simian virus 40 gained notoriety in the 1960s because it was found to be a contaminant of polio and adenovirus vaccines that had been administered to millions of healthy individuals worldwide. The public health implications of this revelation provided the initial impetus for an in-depth study of SV40 biology. Later work showed that SV40 DNA sequences as well as infectious virus are in fact found in human tumors and may have contributed to oncog- esis. It also turned out that SV40 uses mostly cellular machinery to carry out many steps in viral infection, which makes it a powerful probe for examining many fundamental questions in eukaryotic molecular biology. SV40 Pro- cols consolidates a number of well-tested step-by-step techniques in one v- ume; experts with hands-on experience in particular methods give detailed accounts of their optimized experimental protocols, so that the beginner, as well as more experienced researchers, may readily overcome problems of ambiguity often present in the literature. As with other DNA tumor viruses, the response of cultured cells to SV40 infection depends upon the species being infected. Monkey cells s- port virus production, which leads to their death, whereas rodent cells p- duce only the early proteins and acquire a transformed phenotype. Thus, SV40 Protocols is organized in two sections. The first relates to assays of the lytic cycle of the virus, and the second deals with transformation.

Spis treści

Propagation of Wild-Type and Mutant SV40 James D. Tremblay, Kris F. Sachsenmeier, and James M. Pipas Propagation and Assay of JC Virus Christine K. Liu and Walter J. Atwood Propagation and Assay of BK Virus Wendy A. Knowles Molecular Methods for Identification and Genotyping of BK Virus Li Jin Methods for Studying Interactions between Simian Virus 40 T-Antigen and the Viral Origin of Replication Kodangatil R. Sreekumar, Brett A. Barbaro, Andrea E. Prack, and Peter A. Bullock Cell-Free Transcription of SV40 Richard J. Kraus and Janet E. Mertz Quantitation and Structural Analysis of SV40 RNAs Richard J. Kraus and Janet E. Mertz Generation of Recombinant SV40 Vectors for Gene Transfer David S. Strayer, Maria Lamothe, Danlan Wei, Joseph Milano, and Rumi Kondo Production of SV40 Proteins in Insect Cells and In Vitro Packaging of Virions and Pseudovirions Ziv Sandalon and Ariella Oppenheim Using Retroviral Vectors to Express SV40 Tumor Antigens Van Cherington and Cynthia Higgins Neoplastic Transformation Assays Leda Raptis and Adina Vultur Methods to Assess Inhibition of Adipocyte Differentiation by SV40 Large T Antigen Cynthia Higgins and Van Cherington SV40-Mediated Immortalization Mary Judith Tevethia and Harvey L. Ozer Biology of p53 and SV40 Large T Association Ximena Montano Binding of SV40 Large T Antigen to the Retinoblastoma Susceptibility Gene Product and Related Proteins Juan Zalvide, James A. DeCaprio, and Hilde Stubdal. In Sit Electroporation for the Measurement of c-Ras Activation by SVLT Heather L. Brownell and Leda Raptis Role of SV40 Small t in Cell Lysis, Transformation, and Signaling Ximena Montano and Kathy Rundell Cytotoxic T Lymphocytes in SV40 Infections Todd D. Schell and Satvir S. Tevethia Detection of SV40 DNA Sequences in Human Tissue John A. Lednicky and Robert L. Garcea Guide to Techniques in Creating Transgenic Mouse Models Using SV40 T Antigen Ioanna G. Maroulakou and Robin C. Muise-Helmericks Appendix. Index.

Blurb

Extensive research has shown that Simian Virus 40, a contaminant of polio and adenovirus vaccines that may be implicated in human cancers, can also serve as a powerful probe for examining many fundamental questions in molecular biology. In SV40 Protocols, Leda Raptis and a panel of highly experienced investigators describe in step-by-step fashion key techniques for experimentally detecting SV40 in human tumors, for exploiting its use in human gene therapy, and for studying its replication and its mechanisms of neoplastic transformation. Included are methods for growing SV40 and its related viruses in tissue culture, for in vivo and in vitro replication and transcription of SV40 DNA, for the use of retroviral vectors to express SV40 tumor antigens in cultured cells, and for transgenic mouse models based on the SV40 large T antigen. All methods have been optimized for experimental success, and the authors provide cogent discussions of the problems and pitfalls that may be encountered, as well as valuable troubleshooting advice. An appendix lists all companies whose products are cited in the text and includes an Internet directory for locating other reagent sources.
Detailed and highly practical, SV40 Protocols offers both clinical and basic researchers powerful, well-tested tools for research on SV40 replication and neoplastic transformation, as well as techniques for its detection in human tumors and for creating and using powerful new gene therapy vectors.