MemStar: A one‐shot Escherichia coli‐based approach for high‐level bacterial membrane protein production

摘要

OptimisingmembraneproteinproductionyieldsinEscherichiacolicanbetime‐andresource‐consuming.Here,wepresentasimpleandeffectiveMembraneproteinSingleshotamplificationrecipe:MemStar.Thisone‐shotamplificationrecipeisbasedontheE.colistrainLemo21(DE3),thePASM‐5052auto‐inductionmediumand,contradictorily,anIPTGinductionstep.UsingMemStar,productionyieldsformostbacterialmembraneproteinstestedwereimprovedtoreachanaverageof5mgL?1perOD600unit,whichissignificantlyhigherthanyieldsobtainedwithothercommonproductionstrategies.WithMemStar,wehavebeenabletoobtainnewstructuralinformationforseveraltransporters,includingthesodium/protonantiporterNapA...