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dc.contributor.authorMoura, Danielle MN
dc.contributor.authorde Melo Neto, Osvaldo P
dc.contributor.authorCarrington, Mark
dc.date.accessioned2018-12-18T00:32:00Z
dc.date.available2018-12-18T00:32:00Z
dc.date.issued2019-01
dc.identifier.issn0166-6851
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/287078
dc.description.abstractA Trypanosoma brucei cell line is described that produces a visual readout of proteasome activity. The cell line contains an integrated transgene encoding an ubiquitin-green fluorescent protein (GFP) fusion polypeptide responsive to the addition of proteasome inhibitors. A modified version of T. brucei ubiquitin unable to be recognized by deubiquitinases (UbG76V) was fused to eGFP and constitutively expressed. The fusion protein is unstable but addition of the proteasome inhibitor lactacystin stabilizes it and leads to visually detectable GFP. This cell line can be widely used to monitor the efficiency of inhibitor treatment through detection of GFP accumulation in studies involving proteasome-mediated proteolysis, screening of proteasome inhibitors or other events related to the ubiquitin-proteasome pathway.
dc.format.mediumPrint-Electronic
dc.languageeng
dc.publisherElsevier BV
dc.subjectCell Line
dc.subjectHumans
dc.subjectTrypanosoma brucei brucei
dc.subjectTrypanosomiasis, African
dc.subjectProteasome Endopeptidase Complex
dc.subjectGreen Fluorescent Proteins
dc.subjectUbiquitin
dc.subjectTrypanocidal Agents
dc.subjectDrug Evaluation, Preclinical
dc.subjectGenes, Reporter
dc.subjectProteasome Inhibitors
dc.titleA new reporter cell line for studies with proteasome inhibitors in Trypanosoma brucei.
dc.typeArticle
prism.endingPage18
prism.publicationDate2019
prism.publicationNameMol Biochem Parasitol
prism.startingPage15
prism.volume227
dc.identifier.doi10.17863/CAM.34388
dcterms.dateAccepted2018-11-09
rioxxterms.versionofrecord10.1016/j.molbiopara.2018.11.001
rioxxterms.versionAM
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserved
rioxxterms.licenseref.startdate2019-01
dc.contributor.orcidCarrington, Mark [0000-0002-6435-7266]
dc.identifier.eissn1872-9428
rioxxterms.typeJournal Article/Review
pubs.funder-project-idWellcome Trust (085956/Z/08/Z)
rioxxterms.freetoread.startdate2019-11-13


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