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Human Respiratory Syncytial Virus pp 155–164Cite as

Use of Minigenome Systems to Study RSV Transcription

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1442))

Abstract

Minigenome assays have been essential tools in the understanding of viral transcription and RNA replication for respiratory syncytial virus (RSV). Here, we describe the RSV minigenome assay for determining transcription by the viral polymerase in the absence of infection. We detail two different methods of detecting viral RNA synthesis: a firefly luciferase assay for rapid and sensitive measurement of RSV polymerase activity; and a real-time quantitative PCR method for determination of specific effects on the transcription of individual viral genes and the polar transcription gradient of RSV.

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Acknowledgments

The authors thank David Fitzhugh for help in developing the real-time quantitative PCR assay, Martin Moore (Emory University) for the optimized codon expression plasmids for the RSV proteins, Peter Collins (NIAID) for the minigenome and antigenome constructs, and Klaus Conzelmann (Ludwig-Maximillians-Universitat) for the BSR-T7/5 cells. This work was supported in part by a grant from the NIAID and funds from the Joy McCann Culverhouse Endowment.

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Authors and Affiliations

  1. Joy McCann Culverhouse Airway Disease Research Center, Division of Allergy and Immunology, Department of Internal Medicine, University of South Florida Morsani College of Medicine, Tampa, FL, USA

    Michael N. Teng & Kim C. Tran

  2. Department of Internal Medicine, University of South Florida Morsani College of Medicine, 12901 Bruce B. Downs Blvd. MDC 19, Tampa, FL, 336120, USA

    Michael N. Teng

Authors
  1. Michael N. Teng
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  2. Kim C. Tran
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Corresponding author

Correspondence to Michael N. Teng .

Editor information

Editors and Affiliations

  1. College of Veterinary Medicine, University of Georgia, Athens, Georgia, USA

    Ralph A. Tripp

  2. Department of Infectious Diseases, University of Georgia College of Veterinary Medicine, Athens, Georgia, USA

    Patricia A. Jorquera

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Teng, M.N., Tran, K.C. (2016). Use of Minigenome Systems to Study RSV Transcription. In: Tripp, R., Jorquera, P. (eds) Human Respiratory Syncytial Virus. Methods in Molecular Biology, vol 1442. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3687-8_11

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  • DOI: https://doi.org/10.1007/978-1-4939-3687-8_11

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3685-4

  • Online ISBN: 978-1-4939-3687-8

  • eBook Packages: Springer Protocols

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