Abstract
Transcription of the vaccinia virus early genes occurs within the confines of the virion core structure. Therefore, isolated virions are a particularly rich source of proteins that function in early mRNA biosynthesis. Methods are described here for the extraction of purified vaccinia virions to yield protein mixtures with high transcriptional activity on viral early gene templates, responding specifically to both transcriptional initiation and termination signals in the DNA.
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References
Davison, A. J. and Moss, B. (1989) The structure of vaccinia virus early promoters. J. Mol. Biol. 210, 749–769.
Yuen, L. and Moss, B. (1987) Oligonucleotide sequence signaling transcription termination of vaccinia virus early genes. Proc. Nat. Acad. Sci. USA 84, 6417–6421.
Broyles, S. S., Yuen, L., Shuman, S., and Moss, B. (1988) Purification of a factor required for transcription of vaccinia virus early genes. J. Biol. Chem. 263, 10754–10760.
Ahn, B.-Y., Gershon, P. D., and Moss, B. (1994) RNA polymerase associated protein RAP94 confers promoter specificity for initiating transcription of vaccinia virus early genes. J. Biol. Chem. 269, 7552–7557.
Shuman, S., Broyles, S. S., and Moss, B. (1987) Purification and characterization of a transcription termination factor from vaccinia virions. J. Biol. Chem. 262, 12372–12380.
Deng, L. and Shuman, S. (1998) Vaccinia NPH-I, a DExH-box ATPase, is the energy coupling factor for mRNA transcription termination. Genes Dev. 12, 538–546.
Christen, L. M., Sanders, M., Wiler, C., and Niles, E. G. (1998) Vaccinia virus nucleoside triphosphate phosphohydrolase I is an essential viral early gene transcription termination factor. Virology 245, 360–371.
Lackner, C. A. and Condit, R. C. (2000) Vaccinia virus gene A18R DNA helicase is a transcript release factor. J. Biol. Chem. 275, 1485–1494.
Munyon, W. E., Paoletti, E., and Grace, J. J. T. (1967) RNA polymerase activity in purified infectious vaccinia virus. Proc. Nat. Acad. Sci. USA 58, 2280–2288.
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© 2004 Humana Press Inc., Totowa, NJ
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Broyles, S.S., Kremer, M. (2004). An In Vitro Transcription System for Studying Vaccinia Virus Early Genes. In: Isaacs, S.N. (eds) Vaccinia Virus and Poxvirology. Methods in Molecular Biology, vol 269. Humana Press. https://doi.org/10.1385/1-59259-789-0:135
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DOI: https://doi.org/10.1385/1-59259-789-0:135
Publisher Name: Humana Press
Print ISBN: 978-1-58829-229-2
Online ISBN: 978-1-59259-789-5
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