PDF-Proc.Nati.Acad.Sci.USAVol.87,pp.2319-2323,March1990CellBiologyKeratins

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ProcNatlAcadSciUSA871990wereprepared25andtheDNAinsertwasexcisedwithEcoRIisolatedbylowmeltagarosegelelectrophoresisandrandomprimedforexpressionanalysisTheexpressionlevelsandsizesofthemRNA

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Proc.Nati.Acad.Sci.USAVol.87,pp.2319-2323,March1990CellBiologyKeratins: Transcript

ProcNatlAcadSciUSA871990wereprepared25andtheDNAinsertwasexcisedwithEcoRIisolatedbylowmeltagarosegelelectrophoresisandrandomprimedforexpressionanalysisTheexpressionlevelsandsizesofthemRNA. Proc.Natl.Acad.Sci.USA87(1990)6389Corey-Bransona-helixcannotberepresentedwithperfectaccuracyonanylatticeoflowcoordinationnumber,butthehelicesobservedincrystallographydatabasesofrealpro-teinsarelikewis Proc.Natl.Acad.Sci.USA73(1976)29290.6OD0.50.41+mPMU0.3LDEPTH(mm)IIIIIIII00.51.0FIG.1.Atracedcomputeroutputdisplayingtheaveragedopticaldensitiesoflayersinthespecimenofcerebralcortexshownbelow.Thescanni MedicalSciences:Cherif-Zaharetal.Proc.Natl.Acad.Sci.USA87(1990)6245aatcccggcctgcacagagacggacacaggATGAGCTCTAAGTACCCGCGGTCTGTCCGG60MetSerSerLysTyrProArgSerValArg9CGCTGCCTGCCCCTCTGGGCCCTAACACTGGAAGCAGCTC Proc.Natl.Acad.Sci.USA82(1985)855timeswiththesamebufferlackingNP-40andTritonX-100andwereresuspendedinthenick-translationbufferasde-scribedabovecontaining100,Ciof[3H]TTP(117Ci/mmol,NewEnglandNuclear)in Proc.Nat.Acad.Sci.USA70(1978)624~~~~~~~~~~~~24*0aS5!!402040608000020-tO60eo0as--40506~~~~~~~~~~~~~0GROWTHINTHEL00H000)CRW'0NTE0~Z04-20FIG.2.(A)Kineticsofinductionofthechloroplastisoleucyl-tRNAuponexpo Proc.Natl.Acad.Sci.USA93(1996)11529BothLandL*canbeprotonatedonLys-156,butthepKaforprotonationisconsiderablylowerforL*thanforL;pKL*iswhilepKLis10.OneimportanttenetofthismodelisthattheIndsmutationsincre 6148Biochemistry:LittleandHarperTable1.BacterialstrainsemployedSource,reference,StrainRelevantgenotypeorderivationAB1157recA+lexA+sup-37(15)DM49recA+lexA3sup-37(12)DM511recA+IexA3ts1-1sup-37(16)DM1187 Proc.Natl.Acad.Sci.USA90(1993)pHP45QC(13)bySmaIdigestionandgelpurification.pIB415isaderivativeofpIB410(ofIncPreplicon)containingvirAA63/240,avirAderivativelacking178aminoacidsintheperiplasmicdomain(14 5940Biochemistry:Levengoodetal.ProteinPurification.E.coliK17(DE3)cellscontainingpLysSandeitherpSKL17,-18,or-19weregrowntoanOD600of0.5at370Candinducedwith0.4mMIPTGfor2hr.Thecellswereharvested;washedwit Proc.Natl.Acad.Sci.USA82(1985)6415fragmentwasisolatedfroma0.8%agarosegelbyusingtheapparatusforpreparativegelelectrophoresis.DNASequencing.DNAsequencingusingdideoxynucleo-tideswasperformedasdescribedby Proc.Natl.Acad.Sci.USA91(1994)9991pooled;theproteinswereprecipitatedwith(NH4)2SO4anddesaltedasdescribedabove,exceptthattheproteinswereelutedinavolumeof3.5mlusingabuffercontaining50mMNaCland50mMTris-HC Proc.Natl.Acad.Sci.USA82(1985)ofcellsoveriodinevapors.Cellsthataccumulateglycogenstainbrown,whereasthosethatdon'tremainyellow.Quan-titativetrehaloseandglycogendeterminationsweremadeaccordingtoLilliean Proc.Natl.Acad.Sci.USA88(1991)downstreamofthelacpromoterasfollows.PlasmidpHS202(5),whichcontainsnucleotides1-1010(thePvuII-KpnIchromosomefragmentthatencompassesruvA)intheSmaI-KpnIsiteofpUC19,wasdigest 2612Biochemistry:Sancaretal.PstIHaelilt*.JHinfl-TaqIadzJHpa11ti.r4HhaI'recAmRNA-recAprotein-Proc.Nati.Acad.Sci.USA77(1980)EcoRI-9A.-ii4------200020040060080010001200BasepairsFIG.1.Restrictionmapandseq

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