大肠杆菌菌株.docx

大肠杆菌菌株.docx

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大肠杆菌菌株

大肠杆菌菌株

E.coligenotypes

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∙1Nomenclature&Abbreviations

∙2MethylationIssuesinE.coli

∙3Commonlyusedstrains

o3.1AG1

o3.2AB1157

o3.3BL21（AI）

o3.4BL21（DE3）

o3.5BL21（DE3）pLysS

o3.6BNN93

o3.7BW26434,CGSCStrain#7658

o3.8C600

o3.9C600hflA150（Y1073,BNN102）

o3.10D1210

o3.11DB3.1

o3.12DH1

o3.13DH5α

o3.14DH10B（Invitrogen）

o3.15DH12S（Invitrogen）

o3.16DM1（Invitrogen）

o3.17ER2566（NEB）

o3.18ER2267（NEB）

o3.19HB101

o3.20IJ1126

o3.21IJ1127

o3.22JM83

o3.23JM101

o3.24JM103

o3.25JM105

o3.26JM106

o3.27JM107

o3.28JM108

o3.29JM109

o3.30JM109（DE3）

o3.31JM110

o3.32JM2.300

o3.33LE392

o3.34Mach1

o3.35MC1061

o3.36MC4100

o3.37MG1655

o3.38Rosetta（DE3）pLysS

o3.39Rosetta-gami（DE3）pLysS

o3.40RR1

o3.41STBL2（Invitrogen）

o3.42STBL4

o3.43SURE（Stratagene）

o3.44SURE2（Stratagene）

o3.45TOP10（Invitrogen）

o3.46Top10F'（Invitrogen）

o3.47W3110

o3.48XL1-Blue（Stratagene）

o3.49XL2-Blue（Stratagene）

o3.50XL2-BlueMRF'（Stratagene）

o3.51XL1-Red（Stratagene）

o3.52XL10-Gold（Stratagene）

o3.53XL10-GoldKanR（Stratagene）

∙4Othergenotypeinformationsources

∙5References

[edit]Nomenclature&Abbreviations

∙Alistedgenenamemeansthatgenecarriesalossoffunctionmutation,aΔprecedingagenenamemeansthegeneisdeleted.Ifageneisnotlisted,itisnotknowntobemutated.ProphagespresentinwtK-12strains（F,λ,e14,rac）arelistedonlyifabsent.E.coliBstrainsarenaturallylon-anddcm-.

∙F-=DoesnotcarrytheFplasmid

∙F+=CarriestheFplasmid.ThecellisabletomatewithF-throughconjugation.

∙F'[]=CarriesanFplasmidthathashostchromosomalgenesonitfromapreviousrecombinationevent.ThiscellcanalsomatewithF-throughconjugation.ChromosomalgenescarriedintheFplasmidarelistedinbrackets.

∙rB/K+/-=The（B/K）definesthestrainlineage.The+/-indicateswhetherthestrainhasorhasn'tgottherestrictionsystem.

∙mB/K+/-=The（B/K）definesthestrainlineage.The+/-indicateswhetherthestrainhasorhasn'tgotthemodification（methylation）system.

∙hsdS=Bothrestrictionandmethylationofcertainsequencesisdeletedfromthestrain.IfyoutransformDNAfromsuchastrainintoawildtypestrain,itwillbedegraded.

∙hsdR=ForefficienttransformationofclonedunmethylatedDNAfromPCRamplifications

∙INV（）=chromosomalinversionbetweenlocationsindicated

∙ahpC=mutationtoalkylhydroperoxidereductaseconferringdisulfidereductaseactivity

∙ara-14=cannotmetabolizearabinose

∙araD=mutationinL-ribulose-phosphate4-epimeraseblocksarabinosemetabolism

∙cycA=mutationinalaninetransporter;cannotusealanineasacarbonsource

∙dapD=mutationinsuccinyldiaminopimelateaminotransferaseleadstosuccinateor（lysine+methionine）requirement

∙Δ（）=chromosomaldeletionofgenesbetweenthelistedgenes（mayincludeunlistedgenes!

）

∙dam=adeninemethylationatGATCsequencesabolished;highrecombinationefficiency;DNArepairturnedon

∙dcm=cytosinemethylationatsecondCofCCWGGsitesabolished

∙deoR=regulatorygenethatallowsconstitutiveexpressionofdeoxyribosesynthesisgenes;permitsuptakeoflargeplasmids.SeeHanahanD,USPatent4,851,348.

∙dnaJ=oneofthechaparoninsinactivated;stabilizessomemutantproteins

∙dut1=dUTPaseactivityabolished,leadingtoincreaseddUTPconcentrations,allowinguracilinsteadofthymineincorporationinDNA.StableUincorporationrequiresunggenemutationaswell.

∙endA1=ForcleanerpreparationsofDNAandbetterresultsindownstreamapplicationsduetotheeliminationofnon-specificdigestionbyEndonucleaseI

∙（e14）=excisableprophagelikeelementcontainingmcrAgene;presentinK-12butmissinginmanyotherstrains

∙galE=mutantmoreresistanttophageP1infection

∙galK/U=cannotmetabolizegalactose

∙gor=mutationinglutathionereductase;enhancesdisulphidebondformation

∙glnV=suppressionofamber（UAG）stopcodonsbyinsertionofglutamine;requiredforsomephagegrowth

∙gyrA96=mutationinDNAgyrase;conveysnalidixicacidresistance

∙gyrA462=mutationinDNAgyrase;conveysresistancetoccdBcolicingeneproduct

∙hflA150=proteasemutationstabilizingphagecIIprotein;highfrequencyoflysogenizationbyλ

∙Δ（lac）X74=DeletionoftheentirelacoperonaswellassomeflankingDNA.

∙lacIqorlacIQ=overproductionofthelacrepressorprotein;-35siteinpromoterupstreamoflacIismutatedfromGCGCAAtoGTGCAA

∙lacIQ1=overproductionofthelacrepressorprotein;containsa15bpdeletiontocreateoptimal-35siteinpromoterupstreamoflacI

∙lacY=deficientinlactosetransport;deletionoflactosepermease（Mprotein）

∙lacZΔM15=partialdeletionofthelacZgenethatallowsαcomplementationoftheβ-galactosidasegene;requiredforblue/whiteselectiononXGalplates.DeletestheaminoportionoflacZ（aa11-41）.

∙leuB=requiresleucine

∙Δlon=deletionofthelonprotease

∙malA=cannotmetabolizemaltose

∙mcrA=MutationeliminatingrestrictionofDNAmethylatedatthesequenceCmCGG（possiblymCG）.Carriedonthee14prophage（q.v.）

∙mcrB=MutationeliminatingrestrictionofDNAmethylatedatthesequenceRmC

∙metB=requiresmethionine

∙metC=requiresmethionine

∙mrr=MutationeliminatingrestrictionofDNAmethylatedatthesequenceCmAGorGmAC

∙mtlA=cannotmetabilizemannitol

∙（Mu）=Muprophagepresent.Muδmeansthephageisdefective.

∙mutS-mutationinhibitsDNArepairofmismatchesinunmethylatednewlysynthesizedstrands

∙nupG=sameasdeoR

∙ompT=mutationinoutermembraneproteinproteaseVII,reducingproteolysisofexpressedproteins

∙（P1）=CellcarriesaP1prophage.CellsexpresstheP1restrictionsystem.

∙（P2）=CellcarriesaP2prophage.AllowsselectionagainstRed+Gam+λ

∙（φ80）=Cellcarriesthelambdoidprophageφ80.AdefectiveversionofthisphagecarryinglacZM15deletionispresentinsomestrains.

∙pLysS=containspLysSplasmidcarryingchloramphenicolresistanceandphageT7lysozyme,effectiveatattenuatingactivityofT7RNApolymerase,forbetterinhibitionofexpressionundernon-inducedconditions.Thesequencecanbefoundhere.

∙proA/B=requiresproline

∙recA1=ForreducedoccurrenceofunwantedrecombinationinclonedDNA;cellsUVsensitive,deficientinDNArepair

∙recA13=asforrecA1,butinsertslessstable.

∙recBCD=ExonucleaseV;mutationinRecBorRecCreducesgeneralrecombinationbyafactorof100;impairedDNArepair;UVsensitive,easierpropagationofinvertedrepeats

∙recJExonucleaseinvolvedinalternaterecombination

∙relA=relaxedphenotype;permitsRNAsynthesisinabsenceofproteinsynthesis

∙rha=blockedrhamosemetabolism

∙rnc=encodesRnaseIII（rnc-14isacommonnullmutant）

∙rne=encodesRnaseE（rne-3071isacommontemperaturesensitivemutant）

∙rpsL=mutationinribosomalproteinS12conveyingstreptomycinresistance;alsocalledstrA

∙sbcBC=ExoIactivityabolished;usuallypresentinrecBCstrains;recombinationproficient,stableinvertedrepeats

∙srl=cannotmetabolizesorbitol

∙supE=glnV

∙supF=tyrT

∙thi=requiresthiamine

∙thyA=requiresthymidine

∙Tn10=transposonnormallycarryingTetracyclineresistance

∙Tn5=transposonnormallycarryingKanamycinresistance

∙tonA=MutationinoutermembraneproteinconveyingresistancetophageT1andphageT5

∙traD=Mutationeliminatingtransferfactor;preventstransferofFplasmid

∙trxB=mutationinthioredoxinreductase;enhancesdisulphidebondformationinthecytoplasm

∙tsx=outermembraneproteinmutationconveyingresistancetophageT6andcolicinK

∙tyrT=suppressionofamber（UAG）stopcodonsbyinsertionoftyrosine;neededforsomephageinfectionsuchasλgt11.

∙ung1=allowsuraciltoexistinplasmidDNA

∙xyl-5=blockedxylosemetabolism

[edit]MethylationIssuesinE.coli

∙TypeImethylationsystems:

oE.coliK-12restrictsDNAwhichisnotprotectedbyadeninemethylationatsitesAA*C[N6]GTGCorGCA*C[N6]GTT,encodedbythehsdRMSgenes（EcoKI）.Deletionsinthesegenesremoveseithertherestrictionormethylationorbothofthesefunctions.

oE.coliBderivativestrainscontainanhsdRMSsystem（EcoBI）restrictingandprotectiingthesequenceTGA*[N8]TGCTorAGCA*[N8]TCA.

∙ThemcrAgene（carriedonthee14prophage）restrictsDNAwhichismethylatedinCmCWGGormCGsequences（methylationbythedcmgeneproduct）.

∙ThemcrBCgenesrestrictRmCsequences.

∙ThemrrgeneproductrestrictsadeninemethylatedsequencesatCAGorGACsites.

∙E.colimethylatestheadenineinGATC（andthecorrespondingAontheoppositestrand）withthedamgeneproduct.

∙M.EcoKIImethylatesthefirstAatthepalindromicsiteATGCAT（aswellasthecorrespondingAontheoppositestrand）,see（KossykhVG（2004）J.Bact186:

2061-2067PMID15028690）Notethatthisarticlehasbeenretracted;theretractionappearstocenterontextualplagarism,notexperimentalresults.ThehomologytoAvaIIIisreal.IthinkIbelieveit.tk20:

28,9December2005（EST）.RichRobertsreports:

"Wehavetriedourselvestodetectactivitywiththisgeneproductandcannotdetectanymethyltransferaseactivity.InourcaseweusedantibodiesabletodetectN6-methyladenineorN4methylcytosineinDNA.Theoneswehaveareverysensitiveandshouldhavebeenabletodetect5methylgroupsinthewholeE.colichromosome.Nothingwasdetectedinanoverexpressingstrain