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An enhanced toolkit for the generation of knockout and marker-free fluorescent Plasmodium chabaudi
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2020 (Engelska)Ingår i: Wellcome open research, ISSN 2398-502X, Vol. 5, artikel-id 71Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

The rodent parasite Plasmodium chabaudi is an important in vivo model of malaria. The ability to produce chronic infections makes it particularly useful for investigating the development of anti- Plasmodium immunity, as well as features associated with parasite virulence during both the acute and chronic phases of infection. P. chabaudi also undergoes asexual maturation (schizogony) and erythrocyte invasion in culture, so offers an experimentally-amenable in vivo to in vitro model for studying gene function and drug activity during parasite replication. To extend the usefulness of this model, we have further optimised transfection protocols and plasmids for P. chabaudi and generated stable, fluorescent lines that are free from drug-selectable marker genes. These mother-lines show the same infection dynamics as wild-type parasites throughout the lifecycle in mice and mosquitoes; furthermore, their virulence can be increased by serial blood passage and reset by mosquito transmission. We have also adapted the large-insert, linear PlasmoGEM vectors that have revolutionised the scale of experimental genetics in another rodent malaria parasite and used these to generate barcoded P. chabaudi gene-deletion and -tagging vectors for transfection in our fluorescent P. chabaudi mother-lines. This produces a tool-kit of P. chabaudi lines, vectors and transfection approaches that will be of broad utility to the research community.

Ort, förlag, år, upplaga, sidor
Wellcome open research , 2020. Vol. 5, artikel-id 71
Nyckelord [en]
PlasmoGem, Plasmodium chabaudi, Transfection, malaria
Nationell ämneskategori
Mikrobiologi inom det medicinska området
Forskningsämne
mikrobiologi
Identifikatorer
URN: urn:nbn:se:umu:diva-172671DOI: 10.12688/wellcomeopenres.15587.1PubMedID: 32500098Scopus ID: 2-s2.0-85086157248OAI: oai:DiVA.org:umu-172671DiVA, id: diva2:1445436
Tillgänglig från: 2020-06-23 Skapad: 2020-06-23 Senast uppdaterad: 2023-03-23Bibliografiskt granskad

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Billker, OliverBushell, Ellen S.

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Billker, OliverBushell, Ellen S.
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Institutionen för molekylärbiologi (Teknisk-naturvetenskaplig fakultet)Molekylär Infektionsmedicin, Sverige (MIMS)Institutionen för molekylärbiologi (Medicinska fakulteten)
Mikrobiologi inom det medicinska området

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