SV40 Protocols, 2001 Methods in Molecular Biology Series, Vol. 165

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.

Propagation of Wild-Type and Mutant SV40.- Propagation and Assay of the JC Virus.- Propagation and Assay of BK Virus.- Molecular Methods for Identification and Genotyping of BK Virus.- Methods for Studying Interactions Between Simian Virus 40 T-Antigen and the Viral Origin of Replication.- Cell-Free Transcription of SV40.- Quantitation and Structural Analysis of SV40 RNAs.- Generation of Recombinant SV40 Vectors for Gene Transfer.- Production of SV40 Proteins in Insect Cells and In Vitro Packaging of Virions and Pseudovirions.- Using Retroviral Vectors to Express SV40 Tumor Antigens.- Neoplastic Transformation Assays.- Methods to Assess Inhibition of Adipocyte Differentiation by SV40 Large T Antigen.- SV40-Mediated Immortalization.- Biology of p53 and SV40 Large T Association.- Binding of SV40 Large T Antigen to the Retinoblastoma Susceptibility Gene Product and Related Proteins.- In situ Electroporation for the Measurement of c-Ras Activation by SVLT.- Role of SV40 Small t in Cell Lysis, Transformation, and Signaling.- Cytotoxic T Lymphocytes in SV40 Infections.- Detection of SV40 DNA Sequences in Human Tissue.- Guide to Techniques in Creating Transgenic Mouse Models Using SV40 T Antigen.

Includes supplementary material: sn.pub/extras