title: Use of Minigenome Systems to Study RSV Transcription.
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.
authors.affiliation: 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. mteng@health.usf.edu.|Department of Internal Medicine, University of South Florida Morsani College of Medicine, 12901 Bruce B. Downs Blvd. MDC 19, Tampa, FL, 336120, USA. mteng@health.usf.edu.
authors.forename: Michael N
authors.initials: MN
authors.lastname: Teng
authors.affiliation: 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.
authors.forename: Kim C
authors.initials: KC
authors.lastname: Tran
mesh_terms: D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D016679:Genome, Viral; D006801:Humans; D012333:RNA, Messenger / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism; D018113:Respiratory Syncytial Virus, Human / Q000235:genetics / Q000502:physiology*; D059386:Reverse Genetics / Q000379:methods*; D014158:Transcription, Genetic; D014779:Virus Replication
chemical_list: D012333:RNA, Messenger; D012367:RNA, Viral; D015500:Chloramphenicol O-Acetyltransferase
keywords: Luciferase assay; Minigenome; Polymerase; RSV transcription; Real-time quantitative PCR assay