Samenvatting van het vak Moleculaire Biologie uit de opleiding life sciences, ook wel biologie en medisch laboratoriumonderzoek. Per onderwerp zijn de leerdoelen uitgewerkt aan de hand van het boek ''Molecular biology of the cell'' en ''Campbell'', de powerpoints, de extra lesstof en de opdrachten....
Summary Molecular Biology of the Cell 2 (book) (WBFA007-04)
Summary Regenerative Medicine
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Hogeschool Utrecht (HU)
Biologie en Medisch Laboratoriumonderzoek
Moleculaire Biologie (TLSCMOLBIO3V20_2020)
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Theorie .................................................................................................................................................... 4
Thema 1 ................................................................................................................................................... 4
Structuur van het DNA & RNA ................................................................................................................. 4
Alberts pagina 2-7, 100-101, 175-178 ............................................................................................. 4
Campbell 16.1 .................................................................................................................................. 4
DNA Replicatie ......................................................................................................................................... 5
Alberts pagina 239-258 ................................................................................................................... 5
Campbell 16.2 .................................................................................................................................. 5
Endosymbiosetheorie ............................................................................................................................ 11
Alberts pagina 25-29 ..................................................................................................................... 11
Campbell 7.5 .................................................................................................................................. 11
Organisatie van het genoom in de pro- en eukaryoot .......................................................................... 12
Alberts pagina 22-23, 25-29 .......................................................................................................... 12
Campbell 7.2 .................................................................................................................................. 12
Chromosomen ....................................................................................................................................... 12
Alberts pagina 180-193 ................................................................................................................. 12
Alberts pagina 262-265 ................................................................................................................. 12
Campbell 13.2 ................................................................................................................................ 12
DNA condensatie ................................................................................................................................... 14
Alberts pagina 187-193 ................................................................................................................. 14
Thema 2 ................................................................................................................................................. 18
Subtypen RNA en de verschillende functies van deze subtypen .......................................................... 18
Alberts pagina 301-330 ................................................................................................................. 18
Centrale dogma van de moleculaire biologie (van gen naar eiwit)....................................................... 19
Alberts pagina 2-7 ......................................................................................................................... 19
Alberts pagina 301-330 ................................................................................................................. 19
Campbell 17.1 ................................................................................................................................ 19
Transcriptie initiatie, elongatie en terminatie in pro- en eukaryoot .................................................... 20
Alberts pagina 301-330 ................................................................................................................. 20
Campbell 17.2 ................................................................................................................................ 20
RNA-verwerking eukaryoot ................................................................................................................... 24
Alberts pagina 301-330 ................................................................................................................. 24
Campbell 17.3 ................................................................................................................................ 24
Transport van kern naar cytoplasma..................................................................................................... 27
mRNA afbraak prokaryoot..................................................................................................................... 28
1
,mRNA afbraak eukaryoot ...................................................................................................................... 28
Alberts pagina 351-353 ................................................................................................................. 28
Genexpressie regulatie in het algemeen ............................................................................................... 30
Alberts pagina 369-386 ................................................................................................................. 30
Regulatie transcriptie prokaryoot ......................................................................................................... 30
Alberts pagina 369-386 ................................................................................................................. 30
Regulatie transcriptie eukaryoot ........................................................................................................... 32
Alberts pagina 369-386 ................................................................................................................. 32
Thema 3 ................................................................................................................................................. 34
Translatie initiatie, elongatie en terminatie in eukaryoten en prokaryoten......................................... 34
Alberts pagina 334-361 ................................................................................................................. 34
Campbell 17.4 ................................................................................................................................ 34
Post-translationele modificatie ............................................................................................................. 39
Alberts pagina 353-357 ................................................................................................................. 39
Eiwitten ................................................................................................................................................. 40
Alberts pagina 109-121, 127, 165-166 .......................................................................................... 40
Eiwitafbraak........................................................................................................................................... 42
Alberts pagina 357-361 ................................................................................................................. 42
Regulatie translatie ............................................................................................................................... 43
Thema 4 ................................................................................................................................................. 43
Mutagenese........................................................................................................................................... 43
Alberts pagina 217-218, 237-239 .................................................................................................. 43
(Post-) replicatief herstel van DNA en Pre-replicatief herstel van DNA ........................................ 46
Albertspagina 266-288 .................................................................................................................. 46
Meiose ................................................................................................................................................... 50
Alberts pagina 1004-1005 ............................................................................................................. 50
Technieken ............................................................................................................................................ 51
Alberts pagina 463-471, 473-474, 477-478 ................................................................................... 51
Les 1 PCR ............................................................................................................................................... 51
Les 2 Knippen en plakken ...................................................................................................................... 55
Les 3 Amplificatie, isolatie en validatie van recombinant plasmide ..................................................... 58
Les 4 Validatie van recombinant plasmide ............................................................................................ 59
Les 5 Meten van genexpressie .............................................................................................................. 60
Les 6 Kwantitatieve RT-PCR ................................................................................................................... 61
Alberts pagina 502-503 ................................................................................................................. 61
Les 8 en 9 Reverse genetics ................................................................................................................... 63
2
,Les 11 Mutaties opsporen ..................................................................................................................... 66
Alberts pagina 472......................................................................................................................... 66
Les 12 Reverse gentics........................................................................................................................... 72
Alberts pagina 497-498 ................................................................................................................. 72
Opdrachten............................................................................................................................................ 75
Les 1 ....................................................................................................................................................... 75
Les 2 ....................................................................................................................................................... 75
Les 3 ....................................................................................................................................................... 76
Les 4 ....................................................................................................................................................... 77
Les 5 ....................................................................................................................................................... 77
Les 6 ....................................................................................................................................................... 77
Les 7 ....................................................................................................................................................... 78
Les 8 ....................................................................................................................................................... 78
Les 9 ....................................................................................................................................................... 78
Les 10 ..................................................................................................................................................... 78
Les 11 ..................................................................................................................................................... 79
3
, Theorie
Thema 1
Structuur van het DNA & RNA
Alberts pagina 2-7, 100-101, 175-178
Campbell 16.1
• DNA is zuur, omdat de fosfaatgroep een H+ heeft afgestaan aan de omgeving.
• In een volledige draai van de helix zijn 10,4 lagen van basen paren.
• DNA is een rechtsdraaiende dubbele helix.
• DNA is opgebouwd uit een stikstofhoudende base, deoxyribose en een fosfaatgroep. Hoe
meer fosfaatgroepen, hoe negatiever de lading.
• De base zijn Adenine, Guanine, Cytosine en Thymine. (AT en CG). Bij RNA is de T een U.
• De stikstofbase zitten aan de binnenkant. Dit zijn waterstofbruggen.
• De fosfaat-suiker backbone zit aan de buitenkant. Dit zijn covalente bindingen.
• Van der Waalskrachten tussen de basenparen houden het molecuul bij elkaar.
• Tussen de stikstofhoudende base en deoxyribose zit een glycosidische binding.
• Aan de 5’ en de 3’ zijde zitten phosphodiesterbindingen.
• Bij meerdere fosfaatgroepen zitten phosphoanhydride bindingen.
• Elektrostatische krachten= tussen DNA en eiwitten.
• Adenine en Guanine hebben 2 organische ringen. Cytosine en Thymine 1.
• Adenine en Thymine vormen 2 waterstofbindingen. Cytosine en Guanine vormen er 3
(sterker).
• Hoe meer CG, hoe hoger de temperatuur om de bindingen te verbreken, doordat CG 3
waterstofbruggen heeft en AT 2.
• Nucleoside= suiker + base
Nucleotide= nucleoside + 1 of meer fosfaatgroepen.
• Het percentage purines en pyrimidines is altijd gelijk in het genoom.
4
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