Inhoudsopgave
DNA-Modificatie.....................................................................................................................................4
Chapter 1: Studying Genomes................................................................................................................4
Genomen, transcriptomen en proteomen (Genomes, Transcriptomes, and proteomes)..................4
1.1 DNA..............................................................................................................................................5
Genen zijn gemaakt van DNA (genes are made of DNA)................................................................5
DNA is een polymeer van nucleotiden (DNA is a polymer of nucleotides).....................................5
De dubbele helix wordt gestabiliseerd door baseparing en base stacking (The double helix is
stabilized by base pairing and base stacking).................................................................................7
De dubbele helix heeft structurele flexibiliteit (The double helix has structural flexibility)............8
1.2 RNA and the transcriptome..........................................................................................................9
RNA is een tweede type polynucleotide (RNA is a second type of polynucleotide)........................9
Het RNA gehalte van de cel (The RNA content of the cell).............................................................9
Veel RNAs worden gesynthetiseerd als voorlopermoleculen (Many RNAs are synthesized as
precursor molecules)....................................................................................................................10
1.3 Proteins and the proteome.........................................................................................................11
Er zijn 4 hiërarchische niveaus van eiwitstructuur (There are four hierarchical levels of protein
structure)......................................................................................................................................11
Aminozuurdiversiteit ligt ten grondslag aan eiwitdiversiteit (Amino acid diversity underlies
protein diversity)..........................................................................................................................13
De link tussen het transcriptoom en het proteoom (The link between the transcriptome and the
proteome).....................................................................................................................................13
De genetische code is niet universeel (The genetic code is not universal)...................................14
De link tussen een proteoom en de biochemie van de cel (The link between the proteome and
the biochemistry of the cell).........................................................................................................14
Chapter 2: Studying DNA......................................................................................................................16
2.1 Enzymes for DNA manipulation..................................................................................................16
Het werkingsmechanisme van een template afhankelijke DNA polymerase (The mode of action
of a template dependent DNA polymerase).................................................................................17
De typen van DNA polymerase gebruikt in onderzoek (The types of DNA polymerase used in
research).......................................................................................................................................18
Polymerase...................................................................................................................................19
Description...................................................................................................................................19
Main uses.....................................................................................................................................19
DNA polymerase I.........................................................................................................................19
Unmodified E.coli enzyme............................................................................................................19
DNA labeling.................................................................................................................................19
, Klenow polymerase......................................................................................................................19
Modified version of E.coli DNA polymerase I................................................................................19
DNA labelling, chain termination DNA sequencing.......................................................................19
Taq polymerase............................................................................................................................19
Thermus aquaticus DNA polymerase I..........................................................................................19
PCR...............................................................................................................................................19
Reverse transcriptase...................................................................................................................19
RNA-dependent DNA polymerase, obtained from various retroviruses.......................................19
cDNA synthesis.............................................................................................................................19
......................................................................................................................................................19
Nuclease.......................................................................................................................................19
Description...................................................................................................................................19
Main use.......................................................................................................................................19
Restriction endonucleases............................................................................................................19
Sequence-specific DNA endonucleases, from many sources........................................................19
Many applications........................................................................................................................19
S1 nuclease...................................................................................................................................19
Endonuclease specific for single stranded DNA and RNA, from the fungus Aspergillus oryzae....19
Transcript mapping.......................................................................................................................19
Deoxyribonuclease I.....................................................................................................................19
Endonuclease specific for double stranded DNA and RNA from E.coli.........................................19
Nuclease footprinting...................................................................................................................19
Door restrictie-endonucleasen kunnen DNA moleculen op gedefinieerde posities worden
gesneden (Restriction endonucleases enable DNA molecules tob e cut at defined positions).....19
Gelelektroforese wordt gebruikt om de resultaten van een restrictie-digest te onderzoeken (gel
electrophoresis is used to examine the results of a restriction digest).........................................20
Interessante DNA fragmenten kunnen geïdentificeerd worden door een Southern-hybridisatie
(Interesting DNA fragment scan be identified by Southern hybridization)...................................22
Ligasen voegen DNA fragmenten samen (Ligases join DNA fragments together)........................23
Eind modificatie enzymen (End modification enzymes)...............................................................24
2.2 The polymerase chain reaction...................................................................................................24
Een PCR uitvoeren (Carrying out a PCR).......................................................................................24
De snelheid van productvorming kan worden gevolgd tijdens een PCR (The rate of product
formation can be followed during a PCR).....................................................................................25
PCR heeft veel en diverse toepassingen (PCR has many and diverse applications)......................26
2.3 DNA cloning................................................................................................................................27
Waarom is gen klonering belangrijk? (Why is gene cloning important?)......................................27
, De eenvoudigste kloneringsvectoren zijn gebaseerd op E.coli plasmiden (The simplest cloning
vectors are based on E.coli plasmids)...........................................................................................28
Bacteriofagen kunnen ook gebruikt worden als kloneringsvector (Bacteriophages can also be
used as cloning vectors)................................................................................................................29
Vectoren voor langere stukjes DNA (Vectors for longer pieces of DNA).......................................29
DNA gekloneerd in organismen anders dan E.coli (DNA can be cloned in organism other than
E.coli)............................................................................................................................................29
DNA-Modificatie.....................................................................................................................................4
Chapter 1: Studying Genomes................................................................................................................4
Genomen, transcriptomen en proteomen (Genomes, Transcriptomes, and proteomes)..................4
1.1 DNA..............................................................................................................................................5
Genen zijn gemaakt van DNA (genes are made of DNA)................................................................5
DNA is een polymeer van nucleotiden (DNA is a polymer of nucleotides).....................................5
De dubbele helix wordt gestabiliseerd door baseparing en base stacking (The double helix is
stabilized by base pairing and base stacking).................................................................................7
De dubbele helix heeft structurele flexibiliteit (The double helix has structural flexibility)............8
1.2 RNA and the transcriptome..........................................................................................................9
RNA is een tweede type polynucleotide (RNA is a second type of polynucleotide)........................9
Het RNA gehalte van de cel (The RNA content of the cell).............................................................9
Veel RNAs worden gesynthetiseerd als voorlopermoleculen (Many RNAs are synthesized as
precursor molecules)....................................................................................................................10
1.3 Proteins and the proteome.........................................................................................................11
Er zijn 4 hiërarchische niveaus van eiwitstructuur (There are four hierarchical levels of protein
structure)......................................................................................................................................11
Aminozuurdiversiteit ligt ten grondslag aan eiwitdiversiteit (Amino acid diversity underlies
protein diversity)..........................................................................................................................13
De link tussen het transcriptoom en het proteoom (The link between the transcriptome and the
proteome).....................................................................................................................................13
De genetische code is niet universeel (The genetic code is not universal)...................................14
De link tussen een proteoom en de biochemie van de cel (The link between the proteome and
the biochemistry of the cell).........................................................................................................14
Chapter 2: Studying DNA......................................................................................................................16
2.1 Enzymes for DNA manipulation..................................................................................................16
Het werkingsmechanisme van een template afhankelijke DNA polymerase (The mode of action
of a template dependent DNA polymerase).................................................................................17
De typen van DNA polymerase gebruikt in onderzoek (The types of DNA polymerase used in
research).......................................................................................................................................18
Polymerase...................................................................................................................................19
Description...................................................................................................................................19
Main uses.....................................................................................................................................19
DNA polymerase I.........................................................................................................................19
Unmodified E.coli enzyme............................................................................................................19
DNA labeling.................................................................................................................................19
, Klenow polymerase......................................................................................................................19
Modified version of E.coli DNA polymerase I................................................................................19
DNA labelling, chain termination DNA sequencing.......................................................................19
Taq polymerase............................................................................................................................19
Thermus aquaticus DNA polymerase I..........................................................................................19
PCR...............................................................................................................................................19
Reverse transcriptase...................................................................................................................19
RNA-dependent DNA polymerase, obtained from various retroviruses.......................................19
cDNA synthesis.............................................................................................................................19
......................................................................................................................................................19
Nuclease.......................................................................................................................................19
Description...................................................................................................................................19
Main use.......................................................................................................................................19
Restriction endonucleases............................................................................................................19
Sequence-specific DNA endonucleases, from many sources........................................................19
Many applications........................................................................................................................19
S1 nuclease...................................................................................................................................19
Endonuclease specific for single stranded DNA and RNA, from the fungus Aspergillus oryzae....19
Transcript mapping.......................................................................................................................19
Deoxyribonuclease I.....................................................................................................................19
Endonuclease specific for double stranded DNA and RNA from E.coli.........................................19
Nuclease footprinting...................................................................................................................19
Door restrictie-endonucleasen kunnen DNA moleculen op gedefinieerde posities worden
gesneden (Restriction endonucleases enable DNA molecules tob e cut at defined positions).....19
Gelelektroforese wordt gebruikt om de resultaten van een restrictie-digest te onderzoeken (gel
electrophoresis is used to examine the results of a restriction digest).........................................20
Interessante DNA fragmenten kunnen geïdentificeerd worden door een Southern-hybridisatie
(Interesting DNA fragment scan be identified by Southern hybridization)...................................22
Ligasen voegen DNA fragmenten samen (Ligases join DNA fragments together)........................23
Eind modificatie enzymen (End modification enzymes)...............................................................24
2.2 The polymerase chain reaction...................................................................................................24
Een PCR uitvoeren (Carrying out a PCR).......................................................................................24
De snelheid van productvorming kan worden gevolgd tijdens een PCR (The rate of product
formation can be followed during a PCR).....................................................................................25
PCR heeft veel en diverse toepassingen (PCR has many and diverse applications)......................26
2.3 DNA cloning................................................................................................................................27
Waarom is gen klonering belangrijk? (Why is gene cloning important?)......................................27
, De eenvoudigste kloneringsvectoren zijn gebaseerd op E.coli plasmiden (The simplest cloning
vectors are based on E.coli plasmids)...........................................................................................28
Bacteriofagen kunnen ook gebruikt worden als kloneringsvector (Bacteriophages can also be
used as cloning vectors)................................................................................................................29
Vectoren voor langere stukjes DNA (Vectors for longer pieces of DNA).......................................29
DNA gekloneerd in organismen anders dan E.coli (DNA can be cloned in organism other than
E.coli)............................................................................................................................................29
