原创力文档selection of reliable reference genes in caenorhabditis elegans for analysis of nanotoxicity选择可靠的参考基因在线虫nanotoxicity分析.pdfVIP
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selection of reliable reference genes in caenorhabditis elegans for analysis of nanotoxicity选择可靠的参考基因在线虫nanotoxicity分析
Selection of Reliable Reference Genes in Caenorhabditis
elegans for Analysis of Nanotoxicity
Yanqiong Zhang, Dongliang Chen, Michael A. Smith, Baohong Zhang, Xiaoping Pan*
Department of Biology, East Carolina University, Greenville, North Carolina, United States of America
Abstract
Despite rapid development and application of a wide range of manufactured metal oxide nanoparticles (NPs), the
understanding of potential risks of using NPs is less completed, especially at the molecular level. The nematode
Caenorhabditis elegans (C.elegans) has been emerging as an environmental model to study the molecular mechanism of
environmental contaminations, using standard genetic tools such as the real-time quantitative PCR (RT-qPCR). The most
important factor that may affect the accuracy of RT-qPCR is to choose appropriate genes for normalization. In this study, we
selected 13 reference gene candidates (act-1, cdc-42, pmp-3, eif-3.C, actin, act-2, csq-1, Y45F10D.4, tba-1, mdh-1, ama-1,
F35G12.2, and rbd-1) to test their expression stability under different doses of nano-copper oxide (CuO 0, 1, 10, and 50 mg/
mL) using RT-qPCR. Four algorithms, geNorm, NormFinder, BestKeeper, and the comparative DCt method, were employed
to evaluate these 13 candidates expressions. As a result, tba-1, Y45F10D.4 and pmp-3 were the most reliable, which may be
used as reference genes in future study of nanoparticle-induced genetic response using C.elegans.
Citation: Zhang Y, Chen D, Smith MA, Zhang B, Pan X (2012) Selection of Reliable Reference Genes in Caenorhabditis elegans for Analysis of Nanotoxicity. PLoS
ONE 7(3): e31849. doi:10.1371/journal.pone.0031849
Editor: Stephen J. Johnson, University of Kansas, United States of America
Received November 10, 2011; Accepted January 13, 2012; Published March 15, 2012
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