One-Step Purification of Recombinant Human Amelogenin and Use of Amelogenin as a Fusion Partner 英文参考文献.docVIP

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One-Step Purification of Recombinant Human Amelogenin and Use of Amelogenin as a Fusion Partner 英文参考文献.doc

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One-Step Purification of Recombinant Human Amelogenin and Use of Amelogenin as a Fusion Partner 英文参考文献

One-StepPurificationofRecombinantHuman AmelogeninandUseofAmelogeninasaFusionPartner JohanSvenssonBonde,LeifBulow* DepartmentofPureandAppliedBiochemistry,CenterforChemistryandChemicalEngineering,LundUniversity,Lund,Sweden Abstract Amelogenin is an extracellular protein first identified as a matrix component important for formation of dental enamel duringtoothdevelopment.Lately,amelogeninhasalsobeenfoundtohavepositiveeffectsonclinicalimportantareas,such as treatment of periodontal defects, wound healing, and bone regeneration. Here we present a simple method for purification of recombinant human amelogenin expressed in Escherichia coli, based on the solubility properties of amelogenin. The method combines cell lysis with recovery/purification of the protein and generates a .95% pure amelogenininonestepusingintactharvestedcellsasstartingmaterial.Byusingamelogeninasafusionpartnerwecould furtherdemonstratethatthesamemethodalsobe canexplored topurifyothertargetproteins/peptides inaneffective manner. For instance, a fusion between the clinically used protein PTH (parathyroid hormone) and amelogenin was successfullyexpressedandpurified,andtheamelogeninpartcouldberemovedfromPTHbyusingasite-specificprotease. Citation:SvenssonBondeJ,BulowL(2012)One-StepPurificationofRecombinantHumanAmelogeninandUseofAmelogeninasaFusionPartner.PLoSONE7(3): e33269.doi:10.1371/journal.pone.0033269 Editor:AnnaMitraki,UniversityofCrete,Greece ReceivedJanuary9,2012;AcceptedFebruary10,2012;PublishedMarch19,2012 Copyright:?2012SvenssonBondeandBulow.Thisisanopen-accessarticledistributedunderthetermsoftheCreativeCommonsAttributionLicense,which permitsunrestricteduse,distribution,andreproductioninanymedium,providedtheoriginalauthorandsourcearecredited. Funding:ThisstudywassupportedbygrantsfromVinnova(SwedishGovernmentalAgencyforInnovationSystems).Thefunderhadnoroleinstudydesign, datacollectionandanalysis,decisiontopublish,orpreparationofthemanuscript. CompetingInterests:Theauthorshavedeclaredthat