PDF-Proc.Nati.Acad.Sci.USAVol.88,pp.7749-7753,September1991GeneticsSeparat

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ProcNatlAcadSciUSA881991HMRaade21trpl1can100leu23112his31115ura3asdescribed13FourisogenicstrainswereconstructedeachwithadifferentformoftheHMREsilencerwildtypeAEBhmrAAdel35235

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Proc.Nati.Acad.Sci.USAVol.88,pp.7749-7753,September1991GeneticsSeparat: Transcript


ProcNatlAcadSciUSA881991HMRaade21trpl1can100leu23112his31115ura3asdescribed13FourisogenicstrainswereconstructedeachwithadifferentformoftheHMREsilencerwildtypeAEBhmrAAdel35235. Proc.Natl.Acad.Sci.USA87(1990)6389Corey-Bransona-helixcannotberepresentedwithperfectaccuracyonanylatticeoflowcoordinationnumber,butthehelicesobservedincrystallographydatabasesofrealpro-teinsarelikewis Proc.Natl.Acad.Sci.USA87(1990)wereprepared(25),andtheDNAinsertwasexcisedwithEcoRI,isolatedbylowmeltagarosegelelectrophoresis,andrandom-primedforexpressionanalysis.Theexpressionlev-elsandsizesofthemRNA Proc.Natl.Acad.Sci.USA73(1976)29290.6OD0.50.41+mPMU0.3LDEPTH(mm)IIIIIIII00.51.0FIG.1.Atracedcomputeroutputdisplayingtheaveragedopticaldensitiesoflayersinthespecimenofcerebralcortexshownbelow.Thescanni Proc.Nat.Acad.Sci.USA91(1994)9761thepGEM4vector(Promega)forsequencing.Specifically,a1.2-kbPstIfragment,whichhybridizedtothemousecDNAprobe,butnottotalmouseDNA,wassubclonedtogeneratethesequencehomologou 868CellBiology:OsbornandWeberFIGS.1-7.(Legendappearsatbottomofthefollowingpage.)Proc.Nat.Acad.Sci.USA73(1976) Proc.Nat.Acad.Sci.USA73(1976)869FIG.8.Recoveryofcytoplasmicmicrotubulesin3T3cellsafterColc Proc.Natl.Acad.Sci.USA82(1985)855timeswiththesamebufferlackingNP-40andTritonX-100andwereresuspendedinthenick-translationbufferasde-scribedabovecontaining100,Ciof[3H]TTP(117Ci/mmol,NewEnglandNuclear)in Proc.Natl.Acad.Sci.USA93(1996)11529BothLandL*canbeprotonatedonLys-156,butthepKaforprotonationisconsiderablylowerforL*thanforL;pKL*iswhilepKLis10.OneimportanttenetofthismodelisthattheIndsmutationsincre I,fromFlavobacteriumokeanokoites,isamemberoftheunusual,typeIIsclassofrestrictionendonucleasesthatrec-ognizeaspecificDNAsequenceandcleavenonspecificallyashortdistanceawayfromthatsequence(1).Ibindstheco 5940Biochemistry:Levengoodetal.ProteinPurification.E.coliK17(DE3)cellscontainingpLysSandeitherpSKL17,-18,or-19weregrowntoanOD600of0.5at370Candinducedwith0.4mMIPTGfor2hr.Thecellswereharvested;washedwit TheNunproteinofbacteriophageHK022isamemberofthearginine-richmotiffamilyofRNAbindingproteins,whichincludesthephageNtranscriptionantiterminationproteinandtheHIVTatandRevproteins(Fig.1;refs.1±3).Nunbind Proc.Natl.Acad.Sci.USA88(1991)5593Table1.YeaststrainsNuclearMitochondrialStraingenotypegenotypeRef.947aade2ura3-52p09ID41-6aadellysi15161p+15C1036mit-;COXJintron116splicing-defectivepointmutant(trans- Cellsrespondtodevelopmentalcuesandenvironmentalstimulibyalteringgeneexpression.Often,transmissionofdifferentupstreamsignalsinvolvescommoncomponentsyetelicitsdistinct(butappropriate)outcomes.Numerousag Proc.Natl.Acad.Sci.USA93(1996)2583multiplexamplificationasdescribedabove.Theonlydiffer-encebetweenthesetwoproceduresisthatsomePprimersusedinMGSmaycontainsingle-basemismatchestothetemplatestoconvertthe 2612Biochemistry:Sancaretal.PstIHaelilt*.JHinfl-TaqIadzJHpa11ti.r4HhaI'recAmRNA-recAprotein-Proc.Nati.Acad.Sci.USA77(1980)EcoRI-9A.-ii4------200020040060080010001200BasepairsFIG.1.Restrictionmapandseq

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