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Systematic selection of housekeeping genes for gene expression normalization in chicken embryo fibroblasts infected with Newcastle disease virus.
Biochem. Biophys. Res. Commun. 413, 537-540 (2011)
Gene expression analysis is frequently used to analyze the response to viral infection, and 18S RNA, SHDA and GAPDH represent popular house keeping genes (HKGs) often used to normalize gene expression. Here we describe the first systematic selection and evaluation of suitable HKGs for gene expression analysis in chicken embryo fibroblasts (CEF) infected with NDV adapted to the guidelines from Gorzelniak and Ferguson. Our results indicate that ACTB, HPRT1 and HMBS were valuable and stable HKGs, while 18S RNA, GAPDH and SHDA are considerably regulated during the course of infection and thus precluded for normalization. Normalizing the infection dependent gene IFN-a and the infection independent gene B2M to inappropriate HKGs consequently misleads to significant errors in estimating their regulations. Our study emphasizes that even the most popular HKGs like 18S RNA and GAPDH can lead to divergent and inaccurate data interpretation of significant magnitude if not carefully analyzed for stability before.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
Newcastle disease virus; Housekeeping gene; qRT-PCR; Chicken embryo fibroblast; IFN-GAMMA; ALPHA; NDV
Language
english
Publication Year
2011
HGF-reported in Year
2011
ISSN (print) / ISBN
0006-291X
e-ISSN
1090-2104
Quellenangaben
Volume: 413,
Issue: 4,
Pages: 537-540
Publisher
Elsevier
Publishing Place
San Diego, CA
Reviewing status
Peer reviewed
POF-Topic(s)
30202 - Environmental Health
30201 - Metabolic Health
30201 - Metabolic Health
Research field(s)
Lung Research
Genetics and Epidemiology
Genetics and Epidemiology
PSP Element(s)
G-505000-001
G-500600-003
G-500600-003
PubMed ID
21925148
WOS ID
WOS:000295912800008
Scopus ID
80053583557
Erfassungsdatum
2011-11-07