title: Investigation of reference gene expression during human herpesvirus 6B infection indicates peptidylprolyl isomerase A as a stable reference gene and TATA box binding protein as a gene up-regulated by this virus.
abstract: When using relative gene expression for quantification of RNA it is crucial that the reference genes used for normalization do not change with the experimental condition. We aimed at investigating the expressional stability of commonly used reference genes during Human herpesvirus 6B (HHV-6B) infection. Expression of eight commonly used reference genes were investigated with quantitative PCR in a T-cell line infected with HHV-6B. The stability of genes was investigated using the 2(-ΔΔCT) method and the algorithms BestKeeper, GeNorm and NormFinder. Our results indicate that peptidylprolyl isomerase A (PPIA) is the most stably expressed gene while TATA box binding protein (TBP) is the least stably expressed gene during HHV-6B infection. In a confirmatory experiment, TBP was demonstrated to be dose and time dependently upregulated by HHV-6B. The stability of PPIA is in line with other studies investigating different herpesvirus infections whereas the finding that HHV-6B significantly upregulates TBP is novel and most likely specific to HHV-6B.
authors.affiliation: Karolinska Institutet, Department of Clinical Neuroscience, Center for Molecular Medicine L8:00 Stockholm 17176, Sweden. Electronic address: Elin.Engdahl@ki.se.
authors.forename: Elin
authors.initials: E
authors.lastname: Engdahl
authors.affiliation: Karolinska Institutet, Department of Clinical Neuroscience, Center for Molecular Medicine L8:00 Stockholm 17176, Sweden. Electronic address: Nicky.Dunn@ki.se.
authors.forename: Nicky
authors.initials: N
authors.lastname: Dunn
authors.affiliation: Karolinska Institutet, Department of Clinical Neuroscience, Center for Molecular Medicine L8:00 Stockholm 17176, Sweden. Electronic address: Anna.Fogdell-Hahn@ki.se.
authors.forename: Anna
authors.initials: A
authors.lastname: Fogdell-Hahn
mesh_terms: D015415:Biomarkers; D015870:Gene Expression; D015654:Herpesvirus 6, Human / Q000235:genetics*; D006801:Humans; D019696:Peptidylprolyl Isomerase / Q000235:genetics* / Q000378:metabolism; D019349:Roseolovirus Infections / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D035181:TATA-Box Binding Protein / Q000235:genetics* / Q000378:metabolism; D015854:Up-Regulation
chemical_list: D015415:Biomarkers; D035181:TATA-Box Binding Protein; D019696:Peptidylprolyl Isomerase
keywords: Endogenous control gene; HHV-6B; Human herpesvirus 6B; PPIA; Reference gene; TBP