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1、MicrobialBiotechnology(2010)3(4),403411doi:10.1111/j.1751-7915.2009.00148.xMinireviewRevolutionizingmembraneproteinoverexpressioninbacteriaSusanSchlegel,1?MirjamKlepsch,1?2007).ThenaturalabundanceofmembraneproteinsisDimitraGialama,1DavidWickstr?m,1oftentoolo
2、wtoisolatesufficientmaterialforinvitrofunc-DirkJanSlotboom2andJan-WillemdeGier1*tionalandstructuralstudies.Furthermore,theuseof1CenterforBiomembraneResearch,DepartmentofnaturalsourcesexcludesthepossibilityofgeneticallyBiochemistryandBiophysics,StockholmUnive
3、rsity,modifyingproteinstofacilitatetheirdetectionand/orpuri-SE-10691Stockholm,Sweden.?cation,andefficientlylabellingthemforNuclearMag-2DepartmentofBiochemistry,UniversityofGroningen,neticResonance(NMR)andcrystallographicstudies.Nyenborg4,9747AGGroningen,theN
4、etherlands.Twoclassesofmembraneproteinsexist:b-barrelandhelicalbundlemembraneproteins(vonHeijne,1999).b-BarrelmembraneproteinscanoftenbeexpressedinSummaryinclusionbodiesinEscherichiacolifromwhichtheyarereadilyisolatedandrefoldedintheirnativeconformationTheba
5、cteriumEscherichiacoliisthemostwidely(BannwarthandSchulz,2003).Incontrast,forhelicalusedexpressionhostforoverexpressiontrialsofbundlemembraneproteinstheisolationoffunctionalmembraneproteins.Usually,differentstrains,culturematerialfrominclusionbodiesisseldoms
6、uccessful.conditionsandexpressionregimesarescreenedTherefore,helicalbundlemembraneproteinsmustbefortoidentifytheoptimaloverexpressionstrategy.overexpressedinsuchawaythattheyproperlyinsertinHowever,yieldsareoftennotsatisfactory,especiallythemembranefromwhicht
7、heycanbepuri?edafterdeter-foreukaryoticmembraneproteins.Thishasinitiatedagentextractionorwhereintheycanbestudieddirectly.revolutionofmembraneproteinoverexpressioninHere,wewilldealonlywiththeoverexpressionofhelicalbacteria.Recentstudieshaveshownthatitisfeasib
8、lebundlemembraneproteins,hereafterreferredtoasmem-to(i)engineerorselectforE.colistrainswithstronglybraneproteins.improvedmembraneproteinoverexpressioncharac-Escherichiacoliisthemostwidelyusedhos