Samenvatting Moleculaire technieken leerjaar 2 tweede blok
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Course
Moleculaire Technieken
Institution
Hogeschool Arnhem En Nijmegen (HAN)
Deze samenvatting van de moleculaire technieken gaat over de volgende onderwerpen: BL21(DE3)pLysS transformaties, -pBAD/His, inductie, geninformatie (ORI, Fusion proteïn, gen of intrest), pet15b, IPTG, PlysS, cellysis (verschillende methodes), verschillende chromatografieën uitgelegd, westernbl...
-Het effect van de mutaties op de IDH1-enzymactiviteit
-Planning blok1:
>
-Het doel in blok 2:
>De expressie van mutanten te isoleren + purificatie
Made by: Iris Gülcher
, *IDH1 en zijn mutanten tot overexpressie brengen
*Dit om hun enzymactiviteit te analyseren en het effect te bestuderen van de
mutaties
*Lever 500 μg eiwit aan dr. William Leenders voor verder onderzoek
-Planning blok 2:
>
-BL21(DE3)pLysS competente cellen worden gebruikt:
>Deze zijn handig door de combinatie van DE3
*DE gen bevat promoterset van
~PlacUVS
~LacO en
~T7 RNA pol
*
-Belangrijke kenmerken van de plasmiden:
>
Made by: Iris Gülcher
, -BL21(DE3)pLysS transformaties:
>
-pBAD/His:
>De afwezigheid van L-arabinose produceert zeer lage niveaus van transcriptie van PBAD
>In aanwezigheid van L-arabinose, de expressie van Pbad wordt aangezet
-L-arabinose inductie:
>
>
Made by: Iris Gülcher
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