Cell-Specific Monitoring of Protein Synthesis In Vivo 英文参考文献.docVIP

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Cell-Specific Monitoring of Protein Synthesis In Vivo 英文参考文献.doc

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Cell-Specific Monitoring of Protein Synthesis In Vivo 英文参考文献

Cell-SpecificMonitoringofProteinSynthesisInVivo NikosKourtis,NektariosTavernarakis* InstituteofMolecularBiologyandBiotechnology,FoundationforResearchandTechnology-Hellas,Heraklion,Crete,Greece Abstract Analysis of general and specific protein synthesis provides important information, relevant to cellular physiology and function.However,existingmethodologies,involvingmetaboliclabellingbyincorporationofradioactiveaminoacidsinto nascentpolypeptides,cannotbeappliedtomonitorproteinsynthesisinspecificcellsortissues,inlivespecimens.Wehave developed a novel approach for monitoring protein synthesis in specific cells or tissues, in vivo. Fluorescent reporter proteins such as GFP are expressed in specific cells and tissues of interest or throughout animals using appropriate promoters. Protein synthesis rates are assessed by following fluorescence recovery after partial photobleaching of the fluorophoreattargetedsites.Weevaluatethemethodbyexaminingproteinsynthesisratesindiversecelltypesoflive,wild type or mRNA translation-defective Caenorhabditis elegans animals. Because it is non-invasive, our approach allows monitoringofproteinsynthesisinsinglecellsortissueswithintrinsicallydifferentproteinsynthesisrates.Furthermore,it canbereadilyimplementedinotherorganismsorcellculturesystems. Citation:KourtisN,TavernarakisN(2009)Cell-SpecificMonitoringofProteinSynthesisInVivo.PLoSONE4(2):e4547.doi:10.1371/journal.pone.0004547 Editor:MarkR.Cookson,NationalInstitutesofHealth,UnitedStatesofAmerica ReceivedSeptember11,2008;AcceptedJanuary6,2009;PublishedFebruary23,2009 Copyright: ? 2009 Kourtis 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:ThisworkissupportedbygrantsfromEMBO,theEmpeirikeionFoundationandtheEU6thFrameworkProgramme.N.K.isarecipientoftheManasaki scholarship.Thefundershadnoroleinstudydesign,d