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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
biochemistry, genetics and molecular biology
A scalable pipeline for highly effective genetic modification of a malaria parasite
Nature Methods, Volume 8, No. 12, Year 2011
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Description
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. © 2011 Nature America, Inc. All rights reserved.
Authors & Co-Authors
Pfander, Claudia
United Kingdom, Hinxton
Wellcome Sanger Institute
Anar, Burcu
United Kingdom, Hinxton
Wellcome Sanger Institute
Schwach, Frank
United Kingdom, Hinxton
Wellcome Sanger Institute
Otto, Thomas Dan
United Kingdom, Hinxton
Wellcome Sanger Institute
Brochet, M.
United Kingdom, Hinxton
Wellcome Sanger Institute
Volkmann, Katrin
United Kingdom, Hinxton
Wellcome Sanger Institute
Quail, Michael A.
United Kingdom, Hinxton
Wellcome Sanger Institute
Pain, Arnab P.
United Kingdom, Hinxton
Wellcome Sanger Institute
Saudi Arabia, Thuwal
King Abdullah University of Science and Technology
Rosen, Barry S.
United Kingdom, Hinxton
Wellcome Sanger Institute
Skarnes, William C.
United Kingdom, Hinxton
Wellcome Sanger Institute
Rayner, Julian C.
United Kingdom, Hinxton
Wellcome Sanger Institute
Billker, Oliver
United Kingdom, Hinxton
Wellcome Sanger Institute
Statistics
Citations: 104
Authors: 12
Affiliations: 2
Identifiers
Doi:
10.1038/nmeth.1742
ISSN:
15487091
e-ISSN:
15491676
Research Areas
Genetics And Genomics
Infectious Diseases