title: Quantitative Analysis of HSV Gene Expression during Lytic Infection.
abstract: Herpes Simplex Virus (HSV) is a human pathogen that establishes latency and undergoes periodic reactivation, resulting in chronic recurrent lytic infection. HSV lytic infection is characterized by an organized cascade of three gene classes; however, successful transcription and expression of the first, the immediate early class, is critical to the overall success of viral infection. This initial event of lytic infection is also highly dependent on host cell factors. This unit uses RNA interference and small molecule inhibitors to examine the role of host and viral proteins in HSV lytic infection. Methods detailing isolation of viral and host RNA and genomic DNA followed by quantitative real-time PCR allow characterization of impacts on viral transcription and replication, respectively. Western blots can be used to confirm quantitative PCR results. This combination of protocols represents a starting point for researchers interested in virus-host interactions during HSV lytic infection.
authors.affiliation: Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland.
authors.forename: Anne-Marie W
authors.initials: AM
authors.lastname: Turner
authors.forename: Jesse H
authors.initials: JH
authors.lastname: Arbuckle
authors.forename: Thomas M
authors.initials: TM
authors.lastname: Kristie
mesh_terms: D002460:Cell Line; D004279:DNA, Viral / Q000235:genetics; D005347:Fibroblasts / Q000821:virology; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D006801:Humans; D008168:Lung / Q000166:cytology; D034622:RNA Interference; D060888:Real-Time Polymerase Chain Reaction; D018139:Simplexvirus / Q000235:genetics*; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D014776:Virus Cultivation
chemical_list: D004279:DNA, Viral; D014764:Viral Proteins
keywords: Herpes Simplex Virus; RNAi; lytic infection; qPCR; small molecule inhibitors; western blot