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1、Vol456
2、27November2008
3、doi:10.1038/nature07389LETTERSAfast,robustandtunablesyntheticgeneoscillator111,2121,2JesseStricker*,ScottCookson*,MatthewR.Bennett*,WilliamH.Mather,LevS.Tsimring&JeffHastyOnedefininggoalofsyntheticbiologyisthedevelopmentofdecreasespromoteractivity,andthe
4、differentialactivityofthetwo1,13engineering-basedapproachesthatenabletheconstructionoffeedbackloopscandriveoscillatorybehaviour.gene-regulatorynetworksaccordingto‘designspecifications’Theoscillatorcells(denotedJS011)exhibitedubiquitousfluor-1–6escenceoscillationsovertheentire
5、runtimeofeachexperiment(atgeneratedfromcomputationalmodelling.Thisapproachpro-videsasystematicframeworkforexploringhowagivenregulatoryleast4h).Forexample,at0.7%arabinoseand2mMIPTG,morenetworkgeneratesaparticularphenotypicbehaviour.Severalfun-damentalgenecircuitshavebeendevelo
6、pedusingthisapproach,a+Arabinoseb1278–10includingtoggleswitchesandoscillators,andthesehavebeen11appliedinnewcontextssuchastriggeredbiofilmdevelopmentaraCyemGFP812andcellularpopulationcontrol.HerewedescribeanengineeredgeneticoscillatorinEscherichiacolithatisfast,robustandper-−
7、IPTG4sistent,withtunableoscillatoryperiodsasfastas13min.TheoscillatorwasdesignedusingapreviouslymodellednetworkFluorescence(a.u.)0lacI060120180architecturecomprisinglinkedpositiveandnegativefeedbackTime(min)1,13loops.Usingamicrofluidicplatformtailoredforsingle-cellcdmicroscop
8、y,wepreciselycontrolenvironmentalconditionsandmonitoroscillationsinindividualcellsthroughmultiplecycles.Experimentsrevealremarkablerobustnessandpersistenceofoscillationsinthedesignedcircuit;almosteverycellexhibitedlarge-amplitudefluorescenceoscillationsthroughoutobservationru
9、ns.Theoscillatoryperiodcanbetunedbyalteringinducerlevels,temperatureandthemediasource.Computationalmodel-060120180016020180lingdemonstratesthatthekeydesignprincipleforconstructingaefrobustoscillatorisatimedelayinthenegativefeedbackloop,whichcanmechanisticallyarisefromthecasca
10、deofcellularpro-cessesinvolvedinformingafunctionaltranscriptionfacto
