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A scalable pipeline for highly effective genetic modification of a malaria parasite
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2011 (English)In: Nature Methods, ISSN 1548-7091, E-ISSN 1548-7105, Vol. 8, no 12, p. 1078-1082Article in journal (Refereed) Published
Abstract [en]

In malaria parasites, the systematic experimental validation of drug and vaccine targets by reverse genetics is constrained by the inefficiency of homologous recombination and by the difficulty of manipulating adenine and thymine (A+T)-rich DNA of most Plasmodium species in Escherichia coli. We overcame these roadblocks by creating a high-integrity library of Plasmodium berghei genomic DNA (>77% A+T content) in a bacteriophage N15-based vector that can be modified efficiently using the lambda Red method of recombineering. We built a pipeline for generating P. berghei genetic modification vectors at genome scale in serial liquid cultures on 96-well plates. Vectors have long homology arms, which increase recombination frequency up to tenfold over conventional designs. The feasibility of efficient genetic modification at scale will stimulate collaborative, genome-wide knockout and tagging programs for P. berghei.

Place, publisher, year, edition, pages
2011. Vol. 8, no 12, p. 1078-1082
National Category
Cell and Molecular Biology
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URN: urn:nbn:se:umu:diva-165869DOI: 10.1038/nmeth.1742ISI: 000297931700022PubMedID: 22020067OAI: oai:DiVA.org:umu-165869DiVA, id: diva2:1379119
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Wellcome trust, WT089085/Z/09/ZAvailable from: 2019-12-16 Created: 2019-12-16 Last updated: 2019-12-18Bibliographically approved

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Billker, Oliver

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