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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 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 the use of transgenic mouse models based on the SV40 large T antigen. 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.
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Methods in Molecular Biology TM VOLUME 165 SV40 Protocols Edited by Leda Raptis HUMANA PRESS Wild-Type and Mutant SV40 1 1 Propagation of Wild-Type and Mutant SV40 James D. Tremblay, Kris F. Sachsenmeier, and James M. Pipas 1. Introduction Simian virus 40 (SV40) is routinely propagated in established kidney cell lines derived from the African green monkey (1). Generally, one of two cell lines, BSC-1 or CV1, are used for this purpose (2,3). Both of these cell lines are easy to maintain in culture and can be readily frozen and recovered from frozen storage. Wild-type SV40 grows and plaques equally well in both cell lines. Because SV40 uses cellular machinery to carry out many steps in viral infection, it has proved to be a powerful probe of molecular mechanisms of transcription, DNA replication, and growth control. SV40 subverts cellular systems toward virus production by acting on key regulatory cellular targets and pathways. The ana