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thema 2, nucleinezuren en eiwitten

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thema 2, nucleinezuren en eiwitten

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  • December 6, 2018
  • 24
  • 2016/2017
  • Class notes
  • Unknown
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HC: Replicatie en transcriptie
DNA-replicateeisesemi-conservatef
Replicatefoout:eslechutseoneeveere efooutepere 5ebaseparen.
Replicatefooutenewordeneonmiddellijkeverwijderdedoore‘proofreading’eactviuteiutevaneheuteDNA-
polymerase.
DNA-replicateebeeinutebije‘orieinseofereplicatonne(oneeveer
. eineheutemenselijkeeenoom).
Origins of replicaton:eseqoenteevanenocleotdenewaaraan
eeneinitautoreeiwiutezichebinduteomeheuteoiuteelkaareriutsenevanede
dobbeleehelixeuteebeeinnen

Ereiseeeneleadineesutrandeeneeenelaeeineesutrand.eReplicate
verlooputecontnoeeaanedeeleadineesutrandeeneinesutokjeseaanede
laeeineesutrand.
Probleem:eHeuteDNA-polymeraseekanealleeneverleneeneenedos
nieutedeeprimespoutevanedeekeuteneaanleeeen:eheuteheefeeene3n-
OHeeroepenodie.e
Oplossing:ePrimase:esynuthetseeruteeeneRNAe‘primern

MeerdereeenzymenezijnenodieeomeOkazaki fragmenten*eute
synuthetsereneopedeelaeeineesutrand:
- Primase:esynuthetseeruteRNA-primerseopeinutervallenevan
oneeveere2 enocleotden.
- DNA-polymerase:esynutheuthiseeruteDNAeutouteenemeutede
vorieeeRNAeprimer.
- Nuclease:everwijderdedeeprimerseeeeneeauteonutsutaaut.
- Repair DNA polymerase:eVolutedeeeaute(inclodine
proofreadine)
- Ligase:everbindutedeefraemenutenedoorecautalyzerineevan
dooreeeneverbindineemeutedee3nOHeoiuteindeevanedeeene
fraemenutemeutedee5nfosfaauteoiuteindeedeevanedeeanderefraemenute(ATPebenodiede
proces!).
- Helicase:eutrekkeneDNA-sutreneeneoiuteelkaare(ATPevoorenodie)
- Single strand DNA binding proteins:ebindenespecifekeaaneenkelsutreneseDNA.eZoreute
ervooredauteDNAesutreneenenieuteweereuteroeeplakkeneaaneelkaar/dauteheutenieutekapoute
eemaakutekanewordene(doorenocleotases)
- sliding clamp:evoowuteomeDNA,ehooduteDNA-polymeraseevasuteopeDNA.e(bijeOkazakie
fraemenutevaluteslidineeclampevaneDNAeaf,emoeuteopnieoweeemaakuteworden)

*Okazaki fragment:eeemaakuteopelaeeineesutrenee(meerdereeprimersenodieeomeDNAesynuthesee
ineeaneeuteezeten))eeebiedeutosseneprimers.eTossenedeeprimerseworduteheuteDNAeverlenede(2 e
nocleotde)

, Betrokken proteïnen bij
replicate:
- DNAehelicase
- DNAeprimase
- DNAepolymerase
- Enkelsutrenese
DNA-ee
bindineseiwiten
- Slidineeclamp
- Liease
- Noclease




DNA-replicate:ekaneookeineviutroe(koruteefraemenuten)e>ePCR:epolymeraseechainereacton
VerdereePCR-cyclie>eexponenteleeutoenameevaneheuteaanutaleDNAefraemenutenedautedeeexacutee‘utareeutne
seqoenteebevaut
PCR:
 VanekleinesutokjeeDNAeheeleveelemaken)PCR;epolymeraseechainreactoneDNAereplicateeineviutroe
(koruteefraemenuten)e
. eVerhiten->eDNAeoiutelkaar->enkelsutrenes
2. ePrimerse(meutespecifekeeseqoente)ekoelen->eaaneopeDNAezitenee

, 3. Verhiten->DNAesynuthesee(ineaanwezieheidevanenocleotden)e
 cycloseherhalenemeuteeevormdeedobbelsutreneseDNAemolecolene(wanutesynuthesee
eaautenieuteverderedaneprimer)->e2 ecyclie 6emolecolene
Toepassing PCR:e
 Forensischeonderzoek:eDNA fngerprining;eGebaseerdeopewaarnemineedauteheuteeenoomevane
verschillendeeindividoenenieuteexacuteeelijkeis,emaaredauteerebijvoorbeeldeverschilleneaanweziee
zijneine‘repeautnereeions,ezoalsedeeSTRse()eshoruteutandemerepeaut)
 Prenautaleediaenostek
 Deutecteepauthoeenene(ziekuteevaneKlinefeluter;emeerdereeXnen)
 Biomedischeonderzoek


DNA->RNA-synthese= transcripte (overschrijven)

Prokaryoten:especifekeeDNA-voleordese‘verutellenneheuteRNA-polymeraseewaaredeeutranscriptee
moeutebeeinneneenesutoppen:ePROMOTERe(waareutranscripteesutarut)eeneTERMINATOR.

Eukaryoten:e
Promoutor:eeersutebindineevaneandereefacutoren,evoordaute(DNAekanebindeneaanepromoutor)eheute
RNAePol-IIekane‘opsutappenneenemeuteutranscripteekanebeeinnen.eVeeleeenenebeeinnenemeuteeene
TATA-boxe(TAerijkeeebied).eTATAeprouteïneebindutehieraaneeandereeeiwiteneeDNA-
polymeraseekanebindeneaanepromoutoreeeenoeeenocleotdeneaanwezie;esynutheseevindute
plaautse)elanezaameproces,eeebeorutenieutesnel.
Dosevoordautediute(reeolateevaneutranscripteeineeokaryouten)eplaautsvindutezijneactvateeeiwitene
(dieeaaneDNAebinden)enodie,e(medieeruteutranscripteeactvautor;eversnellenevormineebasaleeRNAe
pol-IIeutranscripteecomplex.eHierdoorevinduteeresnellereutranscripteeplaauts->emeereRNA).e

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