版权说明：本文档由用户提供并上传，收益归属内容提供方，若内容存在侵权，请进行举报或认领
文档简介
Chapter14FromGene
toProteinChapter14FromGene
toProteMetabolismTeachesUsAboutGenesMetabolicdefectsstudyingmetabolicdiseasessuggestedthatgenesspecifiedproteinsalkaptonuria(blackurinefromalkaptona.k.a.homogentisicacid)PKU(phenylketonuria)eachdiseaseiscausedby
non-functionalenzymeABCDEGenes
create
phenotype…MetabolismTeachesUsAboutGe2Lecture-034---Protein-Sy教学讲解课件31Gene–1EnzymeHypothesisBeadle&TatumComparedmutantsofbreadmold,NeurosporafunguscreatedmutationsbyX-raytreatmentsX-raysbreakDNAinactivateagenewildtypegrowson“minimal”mediasugars+requiredprecursornutrienttosynthesizeessentialaminoacidsmutantsrequireaddedaminoacidseachtypeofmutantlacksacertainenzymeneededtoproduceacertainaminoacidnon-functionalenzyme=brokengene1Gene–1EnzymeHypothesisBe4Beadle&Tatum1941|1958GeorgeBeadleEdwardTatumBeadle&Tatum1941|1958Geor5Beadle&Tatum’sNeurospora
ExperimentBeadle&Tatum’sNeurosporaEx6So…WhatisaGene?Onegene–oneenzymeallproteinsarecodedbygenesbutnotallproteinsareenzymesOnegene–oneproteineachproteinhasitsowngenebutmanyproteinsarecomposedofseveralpolypeptidesOnegene–onepolypeptidebutmanygenesonlycodeforRNAOnegene–oneproductbutmanygenescodefor
morethanoneproduct…Wheredoesthatleave
us?!So…WhatisaGene?Onegene–7DefiningaGene…“Definingageneisproblematicbecause…onegenecancodeforseveralproteinproducts,somegenescodeonlyforRNA,twogenescanoverlap,andtherearemanyothercomplications.”
–ElizabethPennisi,Science2003genepolypeptide1polypeptide2polypeptide3RNAgeneIt’shardtohuntforwabbits,ifyoudon’tknow
whatawabbit
lookslike.DefiningaGene…“Definingage8proteinRNAThe“CentralDogma”DNAtranscriptiontranslationreplicationHowdowemoveinformationfrom
DNAtoproteins?Forsimplicitysake,let’sgobackto
genesthatcode
forproteins…proteinRNAThe“CentralDogma”D9Fromnucleustocytoplasm…Wherearethegenes?genesareonchromosomesinnucleusWhereareproteinssynthesized?proteinsmadeincytoplasmbyribosomesHowdoestheinformationgetfromnucleustocytoplasm?messengerRNAnucleusFromnucleustocytoplasm…Wher10RNAribosesugarN-basesuracilinsteadofthymineU:AC:GsinglestrandedmRNA,rRNA,tRNA,
siRNA….RNADNAtranscriptionRNAribosesugarRNADNAtranscri11TranscriptionTranscribedDNAstrand=templatestranduntranscribedDNAstrand=codingstrandSynthesisofcomplementaryRNAstrandtranscriptionbubbleEnzymethatfacilitatesthebuildingofRNA:RNApolymeraseTranscriptionTranscribedDNAs12Roleofpromoter1.Wheretostartreading =startingpoint2.Whichstrandtoread =templatestrand3.DirectiononDNA =alwaysreads
DNA
3'5'TranscriptionInitiationRNApolymerasebindstopromotersequenceonDNARoleofpromoterTranscriptionI13TranscriptionElongationRNApolymeraseunwindsDNA
~20basepairsatatimereadsDNA3’5’buildsRNA5’3’(theenzymegovernsthesynthesis!)Noproofreading1error/105basesmanycopiesshortlifenotworthit!TranscriptionElongationNoproo14TranscriptionRNATranscriptionRNA15TranscriptionTerminationRNApolymerasestopsatterminationsequencemRNAleavesnucleusthroughporesRNAGC
hairpinturnTranscriptionTerminationRNAG16Prokaryotevs.EukaryoteGeneticsDifferencesbetweenprokaryotes&eukaryotestime&physicalseparationbetweenprocessesRNAprocessingProkaryotevs.EukaryoteGenet17TranscriptioninEukaryotesonly1prokaryoticenzyme3eukaryoticRNApolymeraseenzymesRNApolymeraseI
onlytranscribesrRNAgenesRNApolymeraseII
transcribesgenesintomRNARNApolymeraseIIIonlytranscribesrRNAgeneseachhasaspecificpromotersequenceitrecognizesTranscriptioninEukaryotesonl18AAAAA3'poly-AtailCH3mRNA5'5'cap3'GPPPEukaryoticPost-transcriptionalProcessingPrimarytranscript
eukaryoticmRNAneedsworkaftertranscriptionProtectmRNAfromRNA-aseenzymesincytoplasmadd5'GcapaddpolyAtailEditoutintronseukaryoticDNAexon=coding(expressed)sequenceintron=noncoding(inbetween)sequenceprimarymRNAtranscriptmaturemRNAtranscriptpre-mRNAsplicedmRNA50-250A’sAAAAA3'poly-AtailCH3mRNA5'5'19PrimaryTranscriptProcessingmRNAprotectingRNAfromRNaseincytoplasmadd5’capaddpolyAtailremoveintronsAUGUGAPrimaryTranscriptProcessingm20ProtectingRNA5’capaddedGtrinucleoside(G-P-P-P)protectsmRNAfromRNase(hydrolyticenzymes)3’poly-Atailadded50-250A’sprotectsmRNAfromRNase(hydrolyticenzymes)helpsexportofRNAfromnucleusUTRUTRProtectingRNA5’capaddedUTRU21Dicing&SplicingmRNAPre-mRNAmRNAeditoutintronsinterveningsequencessplicetogetherexonsexpressedsequencesInhighereukaryotes90%ormoreofgenecanbeintronnooneknowswhy…yetthere’saNobelprizewaiting…“AVERAGE”“gene”=8000bpre-mRNA=8000bmaturemRNA=1200bprotein=400aalotsa“JUNK”!Dicing&SplicingmRNAPre-mRNA22snRNPssmallnuclearRNARNA+proteinsSpliceosomeseveralsnRNPsrecognizesplicesitesequencecut&pasteRNAasribozymesomemRNAcanspliceitselfRNAasenzymeSplicingEnzymessnRNPsSplicingEnzymes23RibozymeSidneyAltmanThomasCech1982|1989YaleUofColoradoRNAasenzymeRibozymeSidneyAltmanThomasCe24SplicingDetailsNoroomformistakes!editing&splicingmustbeexactlyaccurateasinglebaseaddedorlostthrowsoffthereadingframeAUG|CGG|UCC|GAU|AAG|GGC|CAUAUGCGGCUAUGGGUCCGAUAAGGGCCAUAUGCGGUCCGAUAAGGGCCAUAUG|CGG|GUC|CGA|UAA|GGG|CCA|UAUGCGGCUAUGGGUCCGAUAAGGGCCAUAUGCGGGUCCGAUAAGGGCCAUMet|Arg|Ser|Asp
|Lys
|Gly
|His
Met
|Arg|Val
|Arg|STOP|SplicingDetailsNoroomformi25AlternativeSplicingAlternativemRNAsproducedfromsamegenewhenisanintronnotanintron…differentsegmentstreatedasexonsHardtodefine
agene!AlternativeSplicingAlternativ26DiscoveryofSplitGenes1977|1993RichardRobertsPhilipSharpNEBioLabsMITadenoviruscommoncoldDiscoveryofSplitGenes1977|27ssssssssssssssssssssssssssssssssssssStructureofAntibodieslightchainsantigen-bindingsiteheavychainsantigen-bindingsitelightchainlightchainheavychainsBcellmembranevariableregionantigen-bindingsiteYYYYYYYYYYYYYYYYYYYYYYYYYssssssssssssssssssssssssssssss28TranscriptionofgenemRNAchromosomeofundifferentiatedBcellBcellCCDJDNAofdifferentiatedBcellrearrangementofDNAVTranslationofmRNAHowdovertebratesproducemillionsofantibodyproteins,iftheyonlyhaveafewhundredgenescodingforthoseproteins?antibodyByDNArearrangement&somaticmutationvertebratescanproducemillionsof
B&Tcells
TranscriptionofgenemRNAchrom29AAAAAAAAGTP20-30b3'promotertranscriptionstoptranscriptionstartintronsTheTranscriptionalUnit(gene?)transcriptionalunitTACACTDNADNATATA5'RNApolymerasepre-mRNA5'3'translationstarttranslationstopmaturemRNA5'3'UTRUTRexonsenhancer1000+bAAAAAAAAGTP20-30b3'promotertra30Prokaryotevs.EukaryoteGeneticsProkaryotesDNAincytoplasmcircularchromosomenakedDNAnointronsEukaryotesDNAinnucleuslinearchromosomesDNAwoundonhistoneproteinsintronsvs.exonseukaryoticDNAexon=coding(expressed)sequenceintron=noncoding(inbetween)sequenceProkaryotevs.EukaryoteGenet31mRNAFromGenetoProteinDNAtranscriptionnucleuscytoplasmmRNAleavesnucleusthroughnuclearporesproteinssynthesizedbyribosomesusinginstructionsonmRNAaaaaaaaaaaaaaaaaribosomeproteintranslationmRNAFromGenetoProteinDNAtra32TranslationinProkaryotesTranscription&translationaresimultaneousinbacteriaDNAisin
cytoplasmnomRNA
editing
neededTranslationinProkaryotesTran33TACGCACATTTACGTACGCGGDNAAUGCGUGUAAAUGCAUGCGCCmRNAMet
Arg
Val
Asn
Ala
Cys
Alaprotein?Howcanyoucodefor20aminoacidswithonly4nucleotidebases(A,U,G,C)?HowDoesDNACodeforProteinsTACGCACATTTACGTACGCGGDNAAUGCGU34CrackingtheCodeNirenberg&Matthaeidetermined1stcodon–aminoacidmatchUUUcodedforphenylalaninecreatedartificialpoly(U)mRNAaddedmRNAtotesttubeof
ribosomes,tRNA&aminoacidsmRNAsynthesizedsingle
aminoacidpolypeptidechain1960|1968phe–phe–phe–phe–phe–pheCrackingtheCodeNirenberg&M35MarshallNirenbergHeinrichMatthaeiMarshallNirenbergHeinrichMat36TranslationCodonsblocksof3nucleotidesdecodedinto
thesequenceofaminoacidsTranslationCodons37mRNACodesforProteinsinTripletsmRNAAUGCGUGUAAAUGCAUGCGCCmRNAAUGCGUGUAAAUGCAUGCGCCTACGCACATTTACGTACGCGGDNAMetArgValAsnAlaCysAlaprotein?codonribosomemRNAcodesforproteinsintriplets…CODONS!mRNACodesforProteinsinTri38TheCode!ForALLlife!strongestsupportforacommonoriginforalllifeCodeisredundantseveralcodonsforeachaminoacidWhyisthisagoodthing?StartcodonAUGmethionineStopcodonsUGA,UAA,UAGTheCode!ForALLlife!Whyist395'3'3'5'TACGCACATTTACGTACGCGGDNAAUGCGUGUAAAUGCAUGCGCCmRNAanti-codoncodonUACMetGCAArgCAUValamino
acidtRNA3'5'HowareCodonsMatchedtoAminoAcids?5'3'3'5'TACGCACATTTACGTACGCGGD40proteintranscriptioncytoplasmnucleustranslationproteintranscriptioncytoplasmn41tRNAStructure“Cloverleaf”structureanticodonon“cloverleaf”endaminoacidattachedon3'endtRNAStructure“Cloverleaf”st42LoadingtRNAAminoacyltRNAsynthetaseenzymewhichbonds
aminoacidtotRNAendergonicreactionATPAMPenergystoredin
tRNA-aminoacidbondunstablesoitcanreleaseaminoacid
atribosomeLoadingtRNAAminoacyltRNAsy43RibosomesFacilitatecouplingof
tRNAanticodonto
mRNAcodonorganelleorenzyme?StructureribosomalRNA(rRNA)&proteins2subunitslargesmallRibosomesFacilitatecoupling44RibosomesPsite(peptidyl-tRNAsite)holdstRNAcarryinggrowingpolypeptidechainAsite(aminoacyl-tRNAsite)holdstRNAcarryingnextaminoacidtobeaddedtochainEsite(exitsite)emptytRNA
leavesribosome
fromexitsiteRibosomesPsite(peptidyl-tRN45BuildingaPolypeptideInitiationbringstogethermRNA,ribosomesubunits,
proteins&initiatortRNAElongationTerminationBuildingaPolypeptideInitiati46Elongation:GrowingaPolypeptideElongation:GrowingaPolypept47Termination:ReleasePolypeptideReleasefactor“releaseprotein”bondstoAsitebondswatermoleculetopolypeptidechainNowwhathappenstothepolypeptide?Termination:ReleasePolypepti48ProteinTargetingSignalpeptideaddresslabelDestinations:secretionnucleusmitochondriachloroplastscellmembranecytoplasmex.startofasecretorypathwayProteinTargetingSignalpepti49Canyoutelltheeukaryoticstory?DNApre-mRNAribosometRNAamino
acidspolypeptidematuremRNA5'cappolyAtaillargesubunitsmallsubunitaminoacyltRNA
synthetaseEPA5'3'RNApolymeraseexonintrontRNACanyoutelltheeukaryoticst50Puttingitall
together…Don’tforgettheYouTubevideos!Puttingitall
together…Don’t51Chapter14FromGene
toProteinChapter14FromGene
toProteMetabolismTeachesUsAboutGenesMetabolicdefectsstudyingmetabolicdiseasessuggestedthatgenesspecifiedproteinsalkaptonuria(blackurinefromalkaptona.k.a.homogentisicacid)PKU(phenylketonuria)eachdiseaseiscausedby
non-functionalenzymeABCDEGenes
create
phenotype…MetabolismTeachesUsAboutGe53Lecture-034---Protein-Sy教学讲解课件541Gene–1EnzymeHypothesisBeadle&TatumComparedmutantsofbreadmold,NeurosporafunguscreatedmutationsbyX-raytreatmentsX-raysbreakDNAinactivateagenewildtypegrowson“minimal”mediasugars+requiredprecursornutrienttosynthesizeessentialaminoacidsmutantsrequireaddedaminoacidseachtypeofmutantlacksacertainenzymeneededtoproduceacertainaminoacidnon-functionalenzyme=brokengene1Gene–1EnzymeHypothesisBe55Beadle&Tatum1941|1958GeorgeBeadleEdwardTatumBeadle&Tatum1941|1958Geor56Beadle&Tatum’sNeurospora
ExperimentBeadle&Tatum’sNeurosporaEx57So…WhatisaGene?Onegene–oneenzymeallproteinsarecodedbygenesbutnotallproteinsareenzymesOnegene–oneproteineachproteinhasitsowngenebutmanyproteinsarecomposedofseveralpolypeptidesOnegene–onepolypeptidebutmanygenesonlycodeforRNAOnegene–oneproductbutmanygenescodefor
morethanoneproduct…Wheredoesthatleave
us?!So…WhatisaGene?Onegene–58DefiningaGene…“Definingageneisproblematicbecause…onegenecancodeforseveralproteinproducts,somegenescodeonlyforRNA,twogenescanoverlap,andtherearemanyothercomplications.”
–ElizabethPennisi,Science2003genepolypeptide1polypeptide2polypeptide3RNAgeneIt’shardtohuntforwabbits,ifyoudon’tknow
whatawabbit
lookslike.DefiningaGene…“Definingage59proteinRNAThe“CentralDogma”DNAtranscriptiontranslationreplicationHowdowemoveinformationfrom
DNAtoproteins?Forsimplicitysake,let’sgobackto
genesthatcode
forproteins…proteinRNAThe“CentralDogma”D60Fromnucleustocytoplasm…Wherearethegenes?genesareonchromosomesinnucleusWhereareproteinssynthesized?proteinsmadeincytoplasmbyribosomesHowdoestheinformationgetfromnucleustocytoplasm?messengerRNAnucleusFromnucleustocytoplasm…Wher61RNAribosesugarN-basesuracilinsteadofthymineU:AC:GsinglestrandedmRNA,rRNA,tRNA,
siRNA….RNADNAtranscriptionRNAribosesugarRNADNAtranscri62TranscriptionTranscribedDNAstrand=templatestranduntranscribedDNAstrand=codingstrandSynthesisofcomplementaryRNAstrandtranscriptionbubbleEnzymethatfacilitatesthebuildingofRNA:RNApolymeraseTranscriptionTranscribedDNAs63Roleofpromoter1.Wheretostartreading =startingpoint2.Whichstrandtoread =templatestrand3.DirectiononDNA =alwaysreads
DNA
3'5'TranscriptionInitiationRNApolymerasebindstopromotersequenceonDNARoleofpromoterTranscriptionI64TranscriptionElongationRNApolymeraseunwindsDNA
~20basepairsatatimereadsDNA3’5’buildsRNA5’3’(theenzymegovernsthesynthesis!)Noproofreading1error/105basesmanycopiesshortlifenotworthit!TranscriptionElongationNoproo65TranscriptionRNATranscriptionRNA66TranscriptionTerminationRNApolymerasestopsatterminationsequencemRNAleavesnucleusthroughporesRNAGC
hairpinturnTranscriptionTerminationRNAG67Prokaryotevs.EukaryoteGeneticsDifferencesbetweenprokaryotes&eukaryotestime&physicalseparationbetweenprocessesRNAprocessingProkaryotevs.EukaryoteGenet68TranscriptioninEukaryotesonly1prokaryoticenzyme3eukaryoticRNApolymeraseenzymesRNApolymeraseI
onlytranscribesrRNAgenesRNApolymeraseII
transcribesgenesintomRNARNApolymeraseIIIonlytranscribesrRNAgeneseachhasaspecificpromotersequenceitrecognizesTranscriptioninEukaryotesonl69AAAAA3'poly-AtailCH3mRNA5'5'cap3'GPPPEukaryoticPost-transcriptionalProcessingPrimarytranscript
eukaryoticmRNAneedsworkaftertranscriptionProtectmRNAfromRNA-aseenzymesincytoplasmadd5'GcapaddpolyAtailEditoutintronseukaryoticDNAexon=coding(expressed)sequenceintron=noncoding(inbetween)sequenceprimarymRNAtranscriptmaturemRNAtranscriptpre-mRNAsplicedmRNA50-250A’sAAAAA3'poly-AtailCH3mRNA5'5'70PrimaryTranscriptProcessingmRNAprotectingRNAfromRNaseincytoplasmadd5’capaddpolyAtailremoveintronsAUGUGAPrimaryTranscriptProcessingm71ProtectingRNA5’capaddedGtrinucleoside(G-P-P-P)protectsmRNAfromRNase(hydrolyticenzymes)3’poly-Atailadded50-250A’sprotectsmRNAfromRNase(hydrolyticenzymes)helpsexportofRNAfromnucleusUTRUTRProtectingRNA5’capaddedUTRU72Dicing&SplicingmRNAPre-mRNAmRNAeditoutintronsinterveningsequencessplicetogetherexonsexpressedsequencesInhighereukaryotes90%ormoreofgenecanbeintronnooneknowswhy…yetthere’saNobelprizewaiting…“AVERAGE”“gene”=8000bpre-mRNA=8000bmaturemRNA=1200bprotein=400aalotsa“JUNK”!Dicing&SplicingmRNAPre-mRNA73snRNPssmallnuclearRNARNA+proteinsSpliceosomeseveralsnRNPsrecognizesplicesitesequencecut&pasteRNAasribozymesomemRNAcanspliceitselfRNAasenzymeSplicingEnzymessnRNPsSplicingEnzymes74RibozymeSidneyAltmanThomasCech1982|1989YaleUofColoradoRNAasenzymeRibozymeSidneyAltmanThomasCe75SplicingDetailsNoroomformistakes!editing&splicingmustbeexactlyaccurateasinglebaseaddedorlostthrowsoffthereadingframeAUG|CGG|UCC|GAU|AAG|GGC|CAUAUGCGGCUAUGGGUCCGAUAAGGGCCAUAUGCGGUCCGAUAAGGGCCAUAUG|CGG|GUC|CGA|UAA|GGG|CCA|UAUGCGGCUAUGGGUCCGAUAAGGGCCAUAUGCGGGUCCGAUAAGGGCCAUMet|Arg|Ser|Asp
|Lys
|Gly
|His
Met
|Arg|Val
|Arg|STOP|SplicingDetailsNoroomformi76AlternativeSplicingAlternativemRNAsproducedfromsamegenewhenisanintronnotanintron…differentsegmentstreatedasexonsHardtodefine
agene!AlternativeSplicingAlternativ77DiscoveryofSplitGenes1977|1993RichardRobertsPhilipSharpNEBioLabsMITadenoviruscommoncoldDiscoveryofSplitGenes1977|78ssssssssssssssssssssssssssssssssssssStructureofAntibodieslightchainsantigen-bindingsiteheavychainsantigen-bindingsitelightchainlightchainheavychainsBcellmembranevariableregionantigen-bindingsiteYYYYYYYYYYYYYYYYYYYYYYYYYssssssssssssssssssssssssssssss79TranscriptionofgenemRNAchromosomeofundifferentiatedBcellBcellCCDJDNAofdifferentiatedBcellrearrangementofDNAVTranslationofmRNAHowdovertebratesproducemillionsofantibodyproteins,iftheyonlyhaveafewhundredgenescodingforthoseproteins?antibodyByDNArearrangement&somaticmutationvertebratescanproducemillionsof
B&Tcells
TranscriptionofgenemRNAchrom80AAAAAAAAGTP20-30b3'promotertranscriptionstoptranscriptionstartintronsTheTranscriptionalUnit(gene?)transcriptionalunitTACACTDNADNATATA5'RNApolymerasepre-mRNA5'3'translationstarttranslationstopmaturemRNA5'3'UTRUTRexonsenhancer1000+bAAAAAAAAGTP20-30b3'promotertra81Prokaryotevs.EukaryoteGeneticsProkaryotesDNAincytoplasmcircularchromosomenakedDNAnointronsEukaryotesDNAinnucleuslinearchromosomesDNAwoundonhistoneproteinsintronsvs.exonseukaryoticDNAexon=coding(expressed)sequenceintron=noncoding(inbetween)sequenceProkaryotevs.EukaryoteGenet82mRNAFromGenetoProteinDNAtranscriptionnucleuscytoplasmmRNAleavesnucleusthroughnuclearporesproteinssynthesizedbyribosomesusinginstructionsonmRNAaaaaaaaaaaaaaaaaribosomeproteintranslationmRNAFromGenetoProteinDNAtra83TranslationinProkaryotesTranscription&translationaresimultaneousinbacteriaDNAisin
cytoplasmnomRNA
editing
neededTranslationinProkaryotesTran84TACGCACATTTACGTACGCGGDNAAUGCGUGUAAAUGCAUGCGCCmRNAMet
Arg
Val
Asn
Ala
Cys
Alaprotein?Howcanyoucodefor20aminoacidswithonly4nucleotidebases(A,U,G,C)?HowDoesDNACodeforProteinsTACGCACATTTACGTACGCGGDNAAUGCGU85CrackingtheCodeNirenberg&Matthaeidetermined1stcodon–aminoacidmatchUUUcodedforphenylalaninecreatedartificialpoly(U)mRNAaddedmRNAtotesttubeof
ribosomes,tRNA&aminoacidsmRNAsynthesizedsingle
aminoacidpolypeptidechain1960|1968phe–phe–phe–phe–phe–pheCrackingtheCodeNirenberg&M86MarshallNirenbergHeinrichMatthaeiMarshallNirenbergHeinrichMat87TranslationCodonsblocksof3nucleotidesdecodedinto
thesequenceofaminoacidsTranslationCodons88mRNACodesforProteinsinTripletsmRNAAUGCGUGUAAAUGCAUGCGCCmRNAAUGCGUGUAAAUGCAUGCGCCTACGCACATTTACGTACGCGGDNAMetArgValAsnAlaCysAlaprotein?codonribosomemRNAcodesforproteinsintriplets…CODONS!mRNACodesforProteinsinTri89TheCode!ForALLlife
温馨提示
- 1. 本站所有资源如无特殊说明，都需要本地电脑安装OFFICE2007和PDF阅读器。图纸软件为CAD,CAXA,PROE,UG,SolidWorks等.压缩文件请下载最新的WinRAR软件解压。
- 2. 本站的文档不包含任何第三方提供的附件图纸等，如果需要附件，请联系上传者。文件的所有权益归上传用户所有。
- 3. 本站RAR压缩包中若带图纸，网页内容里面会有图纸预览，若没有图纸预览就没有图纸。
- 4. 未经权益所有人同意不得将文件中的内容挪作商业或盈利用途。
- 5. 人人文库网仅提供信息存储空间，仅对用户上传内容的表现方式做保护处理，对用户上传分享的文档内容本身不做任何修改或编辑，并不能对任何下载内容负责。
- 6. 下载文件中如有侵权或不适当内容，请与我们联系，我们立即纠正。
- 7. 本站不保证下载资源的准确性、安全性和完整性, 同时也不承担用户因使用这些下载资源对自己和他人造成任何形式的伤害或损失。
最新文档
- 2026年4月进出口数据解读：断供担忧促使“抢出口”再现
- 烟草种植生物防治技术
- 护理基础：患者出院指导
- 办公室设备维护与故障处理指南
- 网络营销信誉保证承诺书7篇范文
- 实验室检测实力保障承诺函范文4篇
- 护理团队协作：构建高效的护理团队
- 健身教练运动技能培训标准手册
- 校医室护理感染控制与预防
- 牙齿矫正的护理与维护
- 合同授权债权转让协议
- 社会工作探访记录
- 智慧树知道网课《中国古都文化》课后章节测试答案
- 机关宣传稿培训
- 2025年抗肿瘤药物临床合理应用培训试题及答案
- 铁路线路起道作业课件
- 新22G01 砌体房屋结构构造（烧结普通砖、烧结多孔砖）
- DBJ50-T-291-2018 建设工程施工现场安全资料管理标准
- 2025卫生职称(副高)考试小儿内科学高级职称(副高)历年考试真题及答案
- 2025年托育园考试题库及答案
- 民法典与国有企业法治建设专题
评论
0/150
提交评论