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dc.contributor.authorPfander, Claudia
dc.contributor.authorAnar, Burcu
dc.contributor.authorSchwach, Frank
dc.contributor.authorOtto, Thomas D.
dc.contributor.authorBrochet, Mathieu
dc.contributor.authorVolkmann, Katrin
dc.contributor.authorQuail, Michael A.
dc.contributor.authorPain, Arnab
dc.contributor.authorRosen, Barry
dc.contributor.authorSkarnes, William
dc.contributor.authorRayner, Julian C.
dc.contributor.authorBillker, Oliver
dc.date.accessioned2014-08-27T09:49:00Z
dc.date.available2014-08-27T09:49:00Z
dc.date.issued2011-10-23
dc.identifier.citationPfander C, Anar B, Schwach F, Otto TD, Brochet M, et al. (2011) A scalable pipeline for highly effective genetic modification of a malaria parasite. Nature Methods 8: 1078-1082. doi:10.1038/nmeth.1742.
dc.identifier.issn15487091
dc.identifier.pmid22020067
dc.identifier.doi10.1038/nmeth.1742
dc.identifier.urihttp://hdl.handle.net/10754/325358
dc.description.abstractIn 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. © 2011 Nature America, Inc. All rights reserved.
dc.language.isoen
dc.publisherSpringer Nature
dc.rightsUsers may view, print, copy, download and text and data- mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use: http://www.nature.com/authors/editorial_policies/license.html#terms
dc.rightsArchived with thanks to Nature Methods
dc.subjectDNA
dc.subjectbacteriophage
dc.subjectbacteriophage n15
dc.subjectcontrolled study
dc.subjectDNA modification
dc.subjectEscherichia coli
dc.subjectevolutionary homology
dc.subjectgenetic recombination
dc.subjectgenome
dc.subjectPlasmodium berghei
dc.subjectshuttle vector
dc.subjectDNA, Protozoan
dc.subjectDNA, Recombinant
dc.subjectEscherichia coli
dc.subjectGene Library
dc.subjectGenetic Engineering
dc.subjectGenetic Vectors
dc.subjectGenome, Protozoan
dc.subjectHomologous Recombination
dc.subjectMalaria
dc.subjectPlasmodium berghei
dc.subjectEscherichia coli
dc.subjectPlasmodium berghei
dc.titleA scalable pipeline for highly effective genetic modification of a malaria parasite
dc.typeArticle
dc.contributor.departmentBiological and Environmental Sciences and Engineering (BESE) Division
dc.contributor.departmentBioscience Program
dc.contributor.departmentComputational Bioscience Research Center (CBRC)
dc.contributor.departmentPathogen Genomics Laboratory
dc.identifier.journalNature Methods
dc.identifier.pmcidPMC3431185
dc.eprint.versionPost-print
dc.contributor.institutionWellcome Trust Sanger Institute, Hinxton, Cambridge, United Kingdom
dc.contributor.affiliationKing Abdullah University of Science and Technology (KAUST)
kaust.personPain, Arnab
refterms.dateFOA2018-06-13T15:18:43Z
dc.date.published-online2011-10-23
dc.date.published-print2011-12


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