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Samenvatting Moleculaire Biologie: Partim II
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Samenvatting van de cursus Moleculaire Biologie (partim II) van prof. Tavernier en prof. Gerlo.
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Moleculaire BiologiePartim 2: Inleiding tot de
recombinant DNA technologie
1. Principe van kloneren
Kloneren: isoleren ve stof in zuivere toestand gecontroleerd inbrengen v individuele DNA molecule in
autonoom replicerende vector recombinant DNA
Indien stabiel ingebracht in gastheercel kloon ongelimiteerde vermeerdering mogelijk
Essente: recombinant DNA technologie bereid individuele DNA moleculen op grote schaal
Kloneringsaspect & 2 technische pijlers:
Genetsch: inzicht in autonoom replicerende DNA moleculen:
o Plasmiden & virussen & gestheercel (E. Coli)
Biochemisch: karakterisate & zuivering v 2 types enzymes:
o Restricte-enzymes
o DNA ligasen
Structuur v geïsoleerde DNA bepaald dmv DNA sequenteanallse
Human Genome Project
Medische toepassingen:
Therapeutsch:
o Producte v eiwiten
o Gentherapie
Diagnostsch:
o Identicate v mutates aan basis v erfelijke afwijkingen
o Oa PCR
2. Gast/vector systemen
2.1 DNA klonering in plasmiden
Eigenschappen van plasmiden
= circulaire, dubbelstrengige DNA moleculen
Kunnen onafankelijk vh chromosoom bestaan
Symbiotsch verband met gastheercel
Sommige plasmiden bevaten resistenteeenen =voordeel voor gastcel
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