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Selection of reference genes for gene expression studies in rats

Paper ID Volume ID Publish Year Pages File Format Full-Text
23894 43481 2011 10 PDF Available
Title
Selection of reference genes for gene expression studies in rats
Abstract

Selection of the most stable reference gene is critical for a reliable interpretation of gene expression data using RT-PCR. In order so, 17 commonly used genes were analyzed in Wistar rat duodenum, jejunum, ileum and liver following a fat gavage and at two time periods. These reference genes were also tested in liver from Zucker (fa/fa) on a long-term dietary trial. Four strategies were used to select the most suitable reference gene for each tissue: ranking according to biological coefficient of variation and further validation by statistical comparison among groups, geNorm, NormFinder and BestKeeper programs. No agreement was observed among these approaches for a particular gene, nor a common gene for all tissues. Furthermore we demonstrated that normalising using an inadequate reference conveyed into false negative and positive results. The selection of genes provided by BestKeeper resulted in more reliable results than the other statistical packages. According to this program, Tbp, Ubc, Hprt and Rn18s were the best reference genes for duodenum, jejunum, ileum and liver, respectively following a fat gavage in Wistar rats and Rn18s for liver in another rat strain on a long-term dietary intervention. Therefore, BestKeeper is highly recommendable to select the most stable gene to be used as internal standard and the selection of a specific reference expression gene requires a validation for each tissue and experimental design.

Keywords
Actb, β-actin; Arbp, acidic ribosomal phosphoprotein; B2m, β-2-microglobulin; Cck, cholecystokinin; Gapdh, glyceraldehyde-3-phosphate dehydrogenase; Gusb, β-glucuronidase; Hmbs, hidroxymethylbilane synthase; Hprt, hypoxanthine guanine phosphoribosyl trans
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Selection of reference genes for gene expression studies in rats
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 151, Issue 4, 20 February 2011, Pages 325–334
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us