Selection of Reliable Reference Genes in Caenorhabditis elegans for Analysis of Nanotoxicity 英文参考文献.docVIP

SelectionofReliableReferenceGenesinCaenorhabditis elegansforAnalysisofNanotoxicity YanqiongZhang,DongliangChen,MichaelA.Smith,BaohongZhang,XiaopingPan* DepartmentofBiology,EastCarolinaUniversity,Greenville,NorthCarolina,UnitedStatesofAmerica 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 anenvironmental model to study the molecular mechanism of environmental contaminations, using standard genetic tools such as the real-time quantitative PCR (RT-qPCR). The most importantfactorthatmayaffecttheaccuracyofRT-qPCRistochooseappropriategenesfornormalization.Inthisstudy,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,andrbd-1)totesttheirexpressionstabilityunderdifferentdosesofnano-copperoxide(CuO0,1,10,and50 mg/ mL)usingRT-qPCR.Fouralgorithms,geNorm,NormFinder,BestKeeper,andthecomparativeDCtmethod,wereemployed toevaluatethese13candidatesexpressions.Asaresult,tba-1,Y45F10D.4andpmp-3werethemostreliable,whichmaybe usedasreferencegenesinfuturestudyofnanoparticle-inducedgeneticresponseusingC.elegans. Citation:ZhangY,ChenD,SmithMA,ZhangB,PanX(2012)SelectionofReliableReferenceGenesinCaenorhabditiselegansforAnalysisofNanotoxicity.PLoS ONE7(3):e31849.doi:10.1371/journal.pone.0031849 Editor:StephenJ.Johnson,UniversityofKansas,UnitedStatesofAmerica ReceivedNovember10,2011;AcceptedJanuary13,2012;PublishedMarch15,2012 Copyright: ? 2012 Zhang et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:ThisworkwassupportedpartiallybythenewfacultystartupfundofEastCarolinaUniversity,andp