MOLECULAIRE BIOLOGIE
Test jezelf vragen!!!!
HS 5-6: ppt + les belangrijkst !!!! (cursus = basis)
Contents
Hoofdstuk 1 ...................................................................................................................................................................... 7
Grondleggers molec bio ................................................................................................................................................. 7
Inzichten in structuur van erfelijk materiaal ................................................................................................................. 7
model voor structuur van DNA .................................................................................................................................... 7
Centrale dogma ......................................................................................................................................................... 8
Hoofdstuk 2 ...................................................................................................................................................................... 8
Primaire structuur van DNA ............................................................................................................................................ 8
Secundaire structuur ..................................................................................................................................................... 8
Denaturatie en renaturatie van DNA ............................................................................................................................... 9
Tertiaire structuur .......................................................................................................................................................... 9
HS 3: genoomorganisatie en evolutie ................................................................................................................................. 9
genoomsequenties ontrafeld ......................................................................................................................................... 9
variaties in genoomorganisatie ...................................................................................................................................... 10
eukaryote genomen bevatten veel repetitief dna............................................................................................................. 10
extrachromosomaal DNA en horizontale gentransfer ...................................................................................................... 11
genenclusters .............................................................................................................................................................. 12
menselijk genoomproject.............................................................................................................................................. 12
polymorfisme binnen menselijk genoom ........................................................................................................................ 12
genoomseq van hominidae (grote apen) ........................................................................................................................ 13
condensatie van genoom .............................................................................................................................................. 13
hs 4: dna replicatie ........................................................................................................................................................... 13
dna replicatie = semi conservatief (!!!!) .......................................................................................................................... 13
dna replicatie = meestal bidirectioneel .......................................................................................................................... 14
dna polymerasen catalyseren dna synthese van 5’ naar 3’ .............................................................................................. 14
multi-eiwit molec machines verzorgen dna replicatie ..................................................................................................... 14
topo-isomerasen relaxeren supercoiled dna .................................................................................................................. 15
kenmerken eukraryote dna replicatie ............................................................................................................................. 16
laboratoriumtechnieken gebasseerd op DNA replicatie .................................................................................................. 16
sanger sequenering ................................................................................................................................................... 16
pcr (practicum) ......................................................................................................................................................... 16
hs 5: dna schade en herstelmechanismen......................................................................................................................... 21
genetisch materiaal in licht van evolutie ........................................................................................................................ 22
, dna mutaties ................................................................................................................................................................ 22
enu mutagenesis (n-ethyl-N-nitrosourea) ..................................................................................................................... 22
alle mutaties gevolgen? ................................................................................................................................................ 22
theoretisch voorbeeld ................................................................................................................................................... 23
mismatch excisionrepair............................................................................................................................................... 23
base excision repair ...................................................................................................................................................... 24
nucleotide excision repair = NER pathway ...................................................................................................................... 24
herstel T-T dimeer ..................................................................................................................................................... 24
dubbelstrengige breuken .............................................................................................................................................. 24
tp53 ............................................................................................................................................................................. 25
recurrente translocaties ............................................................................................................................................... 25
crispr-cas9................................................................................................................................................................... 25
CRISPR-CAS9-mediated genome editing .................................................................................................................... 26
toepassingen ............................................................................................................................................................ 26
hs 6: ontstaan van genetische variatie ............................................................................................................................... 27
puntmutaties .............................................................................................................................................................. 27
Reverted mozaicism ................................................................................................................................................... 27
Bacteriele resistentie................................................................................................................................................... 27
De novo varianten bij mens ........................................................................................................................................... 27
Somatisch mozaicisme ................................................................................................................................................. 27
genomische herschikkingen .......................................................................................................................................... 27
Homologe recombinatie ............................................................................................................................................ 28
somatische recombinatie: antilichaamdiversiteit ................................................................................................... 28
SMNI1 depletie ........................................................................................................................................................ 29
genduplicatie als gevolg repetitieve seq ..................................................................................................................... 29
hemoglobine genduplicatie – evolutionair voordeel .................................................................................................. 29
transposon .............................................................................................................................................................. 30
retrotransposon ...................................................................................................................................................... 30
samenstelling humaan genoom ................................................................................................................................. 30
retrovirussen ........................................................................................................................................................... 30
carcinogene retrovirussen........................................................................................................................................ 31
rna virussen ............................................................................................................................................................. 31
dna virussen : lytische cyclus .................................................................................................................................. 31
variaties op genoomniveau ........................................................................................................................................... 32
genoom duplicatie in zebravis ....................................................................................................................................... 32
epigenetische variaties ................................................................................................................................................. 32
HS 7: transcriptie en rna maturatie .................................................................................................................................... 32
structuur van rna .......................................................................................................................................................... 32
, boodschapper tussen DNA en eiwit = RNA ..................................................................................................................... 32
basiskenmerken transcriptie ......................................................................................................................................... 33
transcriptie vs replicatie .......................................................................................................................................... 33
transcriptie bij prokaryoten .......................................................................................................................................... 34
promotor ................................................................................................................................................................. 34
rna polymerase ....................................................................................................................................................... 34
regulatie van transcriptie .......................................................................................................................................... 34
terminatie van transcriptie........................................................................................................................................ 35
transcriptie bij eukaryoten ........................................................................................................................................... 35
eukaryoten: 3 verschillende RNA polymerasen .......................................................................................................... 35
eukaryoten CIS-ELEMENTEn ..................................................................................................................................... 35
eukaryote trans-factoren.......................................................................................................................................... 36
DNA condensatie en transcriptionele regulatie ........................................................................................................... 38
transcriptie door rna polymerase I en III en mt transcriptie .......................................................................................... 38
transcriptie terminatie ............................................................................................................................................... 39
centraal dogma ............................................................................................................................................................ 39
rna maturatie ............................................................................................................................................................... 39
5’ capping ................................................................................................................................................................ 39
3’ poly-adenylatie (poly-a) ........................................................................................................................................ 40
splicing .................................................................................................................................................................... 40
mrna editing ................................................................................................................................................................. 42
mrna transport ............................................................................................................................................................. 43
mrna stabiliteit ............................................................................................................................................................. 43
rna interferentie............................................................................................................................................................ 43
RNAi = RNA interferentie ............................................................................................................................................ 43
aso (anti-sense oligonucleotiden) ................................................................................................................................. 45
toepassing ................................................................................................................................................................ 45
rna maturatie ............................................................................................................................................................... 45
hs8: rna translatie ............................................................................................................................................................ 45
informatiestroom ......................................................................................................................................................... 46
eiwitsynthese ............................................................................................................................................................... 46
cracking the code ......................................................................................................................................................... 46
leesraam ..................................................................................................................................................................... 46
ribosoom ..................................................................................................................................................................... 46
polysoom ..................................................................................................................................................................... 46
cry-elektron microscopie .............................................................................................................................................. 46
synthese rrna ............................................................................................................................................................... 47
3 types rna essentieel bij translatie ................................................................................................................................ 47
, eiwitsynthese ............................................................................................................................................................... 47
translatieproces ........................................................................................................................................................... 47
aug – initiatiesignaal .................................................................................................................................................. 47
prokaryote translatie-initiatie ..................................................................................................................................... 48
eukaryote translatie-initiatie ...................................................................................................................................... 48
upstream open reading frames .................................................................................................................................. 49
elongatie .................................................................................................................................................................. 49
terminatie ................................................................................................................................................................. 49
rol van rrna in functie van ribosoom ........................................................................................................................... 49
kwaliteitscontrole ......................................................................................................................................................... 49
prokaryoten: tmRNA ................................................................................................................................................. 49
eukaryoten: PABP-I EIWit .......................................................................................................................................... 50
nonsense mediated decay (NMD) ............................................................................................................................ 50
werking antibiotica/in vitro inhibitie EIWITSYNTHESe ...................................................................................................... 51
toepassing: toxine producerende bacterien ................................................................................................................ 51
geregelde eiwitafbraak .................................................................................................................................................. 51
extracellulair ............................................................................................................................................................ 51
intracellulair ............................................................................................................................................................. 51
foutief opgevouwen eiwitten ziekte ............................................................................................................................ 52
EIWITADDRESSEring ..................................................................................................................................................... 52
sorteren eiwitten aangemaakt op vrije ribosomen ...................................................................................................... 52
sortering naar mt/chloroplasten ............................................................................................................................... 52
import en export van eiwitten naar celkern ................................................................................................................ 52
sortering via ER en golgi ............................................................................................................................................. 53
aantonen rol van signaalpeptide............................................................................................................................... 53
signal recognition particle (SRP)................................................................................................................................. 54
co-translationele translocatie .................................................................................................................................... 54
topogene seq + membr eiwitten ................................................................................................................................. 54
golgi apparaat en vesiculair transport ......................................................................................................................... 55
collageen synthese + secretie .................................................................................................................................... 55
elastine synthese + secretie ...................................................................................................................................... 55
vesiculair transport ................................................................................................................................................... 56
sortering ER resident eiwitten .................................................................................................................................... 56
kwaliteitscontrole in ER ............................................................................................................................................. 57
endocytose en sortering van opgenomen eiwitten ....................................................................................................... 57
transcytose .............................................................................................................................................................. 57
virale infecties via endocytose ................................................................................................................................... 58
adressering van eiwitten voor lysosomale afbraak ...................................................................................................... 58
Test jezelf vragen!!!!
HS 5-6: ppt + les belangrijkst !!!! (cursus = basis)
Contents
Hoofdstuk 1 ...................................................................................................................................................................... 7
Grondleggers molec bio ................................................................................................................................................. 7
Inzichten in structuur van erfelijk materiaal ................................................................................................................. 7
model voor structuur van DNA .................................................................................................................................... 7
Centrale dogma ......................................................................................................................................................... 8
Hoofdstuk 2 ...................................................................................................................................................................... 8
Primaire structuur van DNA ............................................................................................................................................ 8
Secundaire structuur ..................................................................................................................................................... 8
Denaturatie en renaturatie van DNA ............................................................................................................................... 9
Tertiaire structuur .......................................................................................................................................................... 9
HS 3: genoomorganisatie en evolutie ................................................................................................................................. 9
genoomsequenties ontrafeld ......................................................................................................................................... 9
variaties in genoomorganisatie ...................................................................................................................................... 10
eukaryote genomen bevatten veel repetitief dna............................................................................................................. 10
extrachromosomaal DNA en horizontale gentransfer ...................................................................................................... 11
genenclusters .............................................................................................................................................................. 12
menselijk genoomproject.............................................................................................................................................. 12
polymorfisme binnen menselijk genoom ........................................................................................................................ 12
genoomseq van hominidae (grote apen) ........................................................................................................................ 13
condensatie van genoom .............................................................................................................................................. 13
hs 4: dna replicatie ........................................................................................................................................................... 13
dna replicatie = semi conservatief (!!!!) .......................................................................................................................... 13
dna replicatie = meestal bidirectioneel .......................................................................................................................... 14
dna polymerasen catalyseren dna synthese van 5’ naar 3’ .............................................................................................. 14
multi-eiwit molec machines verzorgen dna replicatie ..................................................................................................... 14
topo-isomerasen relaxeren supercoiled dna .................................................................................................................. 15
kenmerken eukraryote dna replicatie ............................................................................................................................. 16
laboratoriumtechnieken gebasseerd op DNA replicatie .................................................................................................. 16
sanger sequenering ................................................................................................................................................... 16
pcr (practicum) ......................................................................................................................................................... 16
hs 5: dna schade en herstelmechanismen......................................................................................................................... 21
genetisch materiaal in licht van evolutie ........................................................................................................................ 22
, dna mutaties ................................................................................................................................................................ 22
enu mutagenesis (n-ethyl-N-nitrosourea) ..................................................................................................................... 22
alle mutaties gevolgen? ................................................................................................................................................ 22
theoretisch voorbeeld ................................................................................................................................................... 23
mismatch excisionrepair............................................................................................................................................... 23
base excision repair ...................................................................................................................................................... 24
nucleotide excision repair = NER pathway ...................................................................................................................... 24
herstel T-T dimeer ..................................................................................................................................................... 24
dubbelstrengige breuken .............................................................................................................................................. 24
tp53 ............................................................................................................................................................................. 25
recurrente translocaties ............................................................................................................................................... 25
crispr-cas9................................................................................................................................................................... 25
CRISPR-CAS9-mediated genome editing .................................................................................................................... 26
toepassingen ............................................................................................................................................................ 26
hs 6: ontstaan van genetische variatie ............................................................................................................................... 27
puntmutaties .............................................................................................................................................................. 27
Reverted mozaicism ................................................................................................................................................... 27
Bacteriele resistentie................................................................................................................................................... 27
De novo varianten bij mens ........................................................................................................................................... 27
Somatisch mozaicisme ................................................................................................................................................. 27
genomische herschikkingen .......................................................................................................................................... 27
Homologe recombinatie ............................................................................................................................................ 28
somatische recombinatie: antilichaamdiversiteit ................................................................................................... 28
SMNI1 depletie ........................................................................................................................................................ 29
genduplicatie als gevolg repetitieve seq ..................................................................................................................... 29
hemoglobine genduplicatie – evolutionair voordeel .................................................................................................. 29
transposon .............................................................................................................................................................. 30
retrotransposon ...................................................................................................................................................... 30
samenstelling humaan genoom ................................................................................................................................. 30
retrovirussen ........................................................................................................................................................... 30
carcinogene retrovirussen........................................................................................................................................ 31
rna virussen ............................................................................................................................................................. 31
dna virussen : lytische cyclus .................................................................................................................................. 31
variaties op genoomniveau ........................................................................................................................................... 32
genoom duplicatie in zebravis ....................................................................................................................................... 32
epigenetische variaties ................................................................................................................................................. 32
HS 7: transcriptie en rna maturatie .................................................................................................................................... 32
structuur van rna .......................................................................................................................................................... 32
, boodschapper tussen DNA en eiwit = RNA ..................................................................................................................... 32
basiskenmerken transcriptie ......................................................................................................................................... 33
transcriptie vs replicatie .......................................................................................................................................... 33
transcriptie bij prokaryoten .......................................................................................................................................... 34
promotor ................................................................................................................................................................. 34
rna polymerase ....................................................................................................................................................... 34
regulatie van transcriptie .......................................................................................................................................... 34
terminatie van transcriptie........................................................................................................................................ 35
transcriptie bij eukaryoten ........................................................................................................................................... 35
eukaryoten: 3 verschillende RNA polymerasen .......................................................................................................... 35
eukaryoten CIS-ELEMENTEn ..................................................................................................................................... 35
eukaryote trans-factoren.......................................................................................................................................... 36
DNA condensatie en transcriptionele regulatie ........................................................................................................... 38
transcriptie door rna polymerase I en III en mt transcriptie .......................................................................................... 38
transcriptie terminatie ............................................................................................................................................... 39
centraal dogma ............................................................................................................................................................ 39
rna maturatie ............................................................................................................................................................... 39
5’ capping ................................................................................................................................................................ 39
3’ poly-adenylatie (poly-a) ........................................................................................................................................ 40
splicing .................................................................................................................................................................... 40
mrna editing ................................................................................................................................................................. 42
mrna transport ............................................................................................................................................................. 43
mrna stabiliteit ............................................................................................................................................................. 43
rna interferentie............................................................................................................................................................ 43
RNAi = RNA interferentie ............................................................................................................................................ 43
aso (anti-sense oligonucleotiden) ................................................................................................................................. 45
toepassing ................................................................................................................................................................ 45
rna maturatie ............................................................................................................................................................... 45
hs8: rna translatie ............................................................................................................................................................ 45
informatiestroom ......................................................................................................................................................... 46
eiwitsynthese ............................................................................................................................................................... 46
cracking the code ......................................................................................................................................................... 46
leesraam ..................................................................................................................................................................... 46
ribosoom ..................................................................................................................................................................... 46
polysoom ..................................................................................................................................................................... 46
cry-elektron microscopie .............................................................................................................................................. 46
synthese rrna ............................................................................................................................................................... 47
3 types rna essentieel bij translatie ................................................................................................................................ 47
, eiwitsynthese ............................................................................................................................................................... 47
translatieproces ........................................................................................................................................................... 47
aug – initiatiesignaal .................................................................................................................................................. 47
prokaryote translatie-initiatie ..................................................................................................................................... 48
eukaryote translatie-initiatie ...................................................................................................................................... 48
upstream open reading frames .................................................................................................................................. 49
elongatie .................................................................................................................................................................. 49
terminatie ................................................................................................................................................................. 49
rol van rrna in functie van ribosoom ........................................................................................................................... 49
kwaliteitscontrole ......................................................................................................................................................... 49
prokaryoten: tmRNA ................................................................................................................................................. 49
eukaryoten: PABP-I EIWit .......................................................................................................................................... 50
nonsense mediated decay (NMD) ............................................................................................................................ 50
werking antibiotica/in vitro inhibitie EIWITSYNTHESe ...................................................................................................... 51
toepassing: toxine producerende bacterien ................................................................................................................ 51
geregelde eiwitafbraak .................................................................................................................................................. 51
extracellulair ............................................................................................................................................................ 51
intracellulair ............................................................................................................................................................. 51
foutief opgevouwen eiwitten ziekte ............................................................................................................................ 52
EIWITADDRESSEring ..................................................................................................................................................... 52
sorteren eiwitten aangemaakt op vrije ribosomen ...................................................................................................... 52
sortering naar mt/chloroplasten ............................................................................................................................... 52
import en export van eiwitten naar celkern ................................................................................................................ 52
sortering via ER en golgi ............................................................................................................................................. 53
aantonen rol van signaalpeptide............................................................................................................................... 53
signal recognition particle (SRP)................................................................................................................................. 54
co-translationele translocatie .................................................................................................................................... 54
topogene seq + membr eiwitten ................................................................................................................................. 54
golgi apparaat en vesiculair transport ......................................................................................................................... 55
collageen synthese + secretie .................................................................................................................................... 55
elastine synthese + secretie ...................................................................................................................................... 55
vesiculair transport ................................................................................................................................................... 56
sortering ER resident eiwitten .................................................................................................................................... 56
kwaliteitscontrole in ER ............................................................................................................................................. 57
endocytose en sortering van opgenomen eiwitten ....................................................................................................... 57
transcytose .............................................................................................................................................................. 57
virale infecties via endocytose ................................................................................................................................... 58
adressering van eiwitten voor lysosomale afbraak ...................................................................................................... 58
