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Expression and analysis of members of the Nudix hydrolase superfamily
KTH, School of Biotechnology (BIO).
2014 (English)Independent thesis Advanced level (degree of Master (Two Years)), 20 credits / 30 HE creditsStudent thesis
Abstract [en]

Inhibition of the Nudix hydrolase MTH1 specifically kills cancer cells. Therefore, it is of interst to purify and characterize other members of the Nudix hydrolase super family in order to find new potential targets for cancer therapy. cDNA encoding seven different Nudix hydrolases, were amplified, cleaved, and ligated into the bacterial pET28 expression vectors followed by transformation into Top10 E.coli cells. Subsequently, the plasmid construct was extracted and sequenzed. Plasmid constructs with correct sequences were transformed into different strains of E.coli for expression of the Nudix hydrolase in question. Out of the seven different Nudix hydrolases expressed, three were managed to be expressed in a soluble form. One of the nudix hydrolases that was expressed in a soluble form, Nudt2, was expressed in a larger scale and was shown to be active. Moreover, it was possible to inhibit Nudt2 with KF, which can be used as a control when screening for inhibitors. 

Place, publisher, year, edition, pages
2014.
Keyword [en]
Nudix, hydrolase, IMAC, Enzyme, Cancer
National Category
Engineering and Technology
Identifiers
URN: urn:nbn:se:kth:diva-163683OAI: oai:DiVA.org:kth-163683DiVA: diva2:801790
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Available from: 2015-04-14 Created: 2015-04-10 Last updated: 2015-09-17Bibliographically approved

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