Bottom-Up Engineering of Biological Systems through Standard Bricks A Modularity Study on Basic Parts and Devices 英文参考文献.docVIP

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Bottom-Up Engineering of Biological Systems through Standard Bricks A Modularity Study on Basic Parts and Devices 英文参考文献.doc

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Bottom-Up Engineering of Biological Systems through Standard Bricks A Modularity Study on Basic Parts and Devices 英文参考文献

Bottom-UpEngineeringofBiologicalSystemsthrough StandardBricks:AModularityStudyonBasicPartsand Devices LorenzoPasotti1,2,Nicolo` Politi1,2,SusannaZucca1,2,MariaGabriellaCusellaDeAngelis2, PaoloMagni1,2 * 1DipartimentodiIngegneriaIndustrialeedell’Informazione,Universita` degliStudidiPavia,Pavia,Italy,2CentrodiIngegneriaTissutale,Universita` degliStudidiPavia, Pavia,Italy Abstract Background:Modularityisacrucialissueintheengineeringworld,asitenablesengineerstoachievepredictableoutcomes whendifferentcomponentsareinterconnected.SyntheticBiologyaimstoapplykeyconceptsofengineeringtodesignand constructnewbiologicalsystemsthatexhibitapredictablebehaviour.Evenifphysicalandmeasurementstandardshave been recently proposed to facilitate the assembly and characterization of biological components, real modularity is still amajorresearchissue.Thesuccessofthebottom-upapproachstrictlydependsonthecleardefinitionofthelimitsinwhich biologicalfunctionscanbepredictable. Results:Themodularityoftranscription-basedbiologicalcomponentshasbeeninvestigatedinseveralconditions.First,the activityofasetofpromoterswasquantifiedinEscherichiacoliviadifferentmeasurementsystems(i.e.,differentplasmids, reportergenes,ribosomebindingsites)relativetoaninvivoreferencepromoter.Second,promoteractivityvariationwas measured when two independent gene expression cassettes were assembled in the same system. Third, the interchangeabilityofinputmodules(asetofconstitutivepromotersandtworegulatedpromoters)connectedtoafixed output device (a logic inverter) expressing GFP was evaluated. The three input modules provide tunable transcriptional signalsthatdrivetheoutputdevice.Ifmodularitypersists,identicaltranscriptionalsignalstriggeridenticalGFPoutputs.To verifythis,alltheinputdeviceswereindividuallycharacterizedandthentheinput-outputcharacteristicofthelogicinverter wasderivedinthedifferentconfigurations. Conclusions:Promotersactivities(referredtoastandardpromoter)canvarywhentheyaremeasuredviadifferentreporter