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Science 2 June 1995:
Vol. 268. no. 5215, pp. 1358 - 1362
DOI: 10.1126/science.7761856
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Articles

Science, Vol 268, Issue 5215, 1358-1362
Copyright © 1995 by American Association for the Advancement of Science

articles

Stable transfection of malaria parasite blood stages

MR van Dijk, AP Waters, and CJ Janse

Department of Parasitology, University of Leiden, Netherlands.

Genetic manipulation of malaria parasites would revolutionize the study of this group of pathogens and have implications for vaccine and drug development. This report describes the stable, drug-selectable genetic transformation of the clinically relevant intracellular blood stages of a malaria parasite. A plasmid transfection vector carrying the gene locus that encodes a drug-resistant form of the bifunctional enzyme dihydrofolate reductase-thymidylate synthase from the rodent malaria parasite Plasmodium berghei was constructed. Derivatives of this vector were introduced into merozoites of P. berghei by electroporation, and parasites were selected for successful transformation in the rodent host on the basis of resistance to pyrimethamine. The plasmids were present in a circular, unrearranged form that replicated episomally to an observed maximum of 15 copies per cell in drug-resistant populations.


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Malaria-Specific and Nonspecific Activation of CD8+ T Cells during Blood Stage of Plasmodium berghei Infection.
M. Miyakoda, D. Kimura, M. Yuda, Y. Chinzei, Y. Shibata, K. Honma, and K. Yui (2008)
J. Immunol. 181, 1420-1428
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Towards systematic identification of Plasmodium essential genes by transposon shuttle mutagenesis.
H. Sakamoto, S. Thiberge, S. Akerman, C. J. Janse, T. G. Carvalho, and R. Menard (2005)
Nucleic Acids Res. 33, e174
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High-efficiency transformation of Plasmodium falciparum by the lepidopteran transposable element piggyBac.
B. Balu, D. A. Shoue, M. J. Fraser Jr., and J. H. Adams (2005)
PNAS 102, 16391-16396
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Mechanisms of Resistance of Malaria Parasites to Antifolates.
A. Gregson and C. V. Plowe (2005)
Pharmacol. Rev. 57, 117-145
   Abstract »    Full Text »    PDF »
Conditional mutagenesis using site-specific recombination in Plasmodium berghei.
T. G. Carvalho, S. Thiberge, H. Sakamoto, and R. Menard (2004)
PNAS 101, 14931-14936
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Discovery of Gene Function by Expression Profiling of the Malaria Parasite Life Cycle.
K. G. Le Roch, Y. Zhou, P. L. Blair, M. Grainger, J. K. Moch, J. D. Haynes, P. De la Vega, A. A. Holder, S. Batalov, D. J. Carucci, et al. (2003)
Science 301, 1503-1508
   Abstract »    Full Text »    PDF »
Transfected Plasmodium knowlesi Produces Bioactive Host Gamma Interferon: a New Perspective for Modulating Immune Responses to Malaria Parasites.
H. Ozwara, J. A. M. Langermans, C. H. M. Kocken, A. van der Wel, P. H. van der Meide, R. A. W. Vervenne, J. M. Mwenda, and A. W. Thomas (2003)
Infect. Immun. 71, 4375-4381
   Abstract »    Full Text »    PDF »
Plasmodium knowlesi Provides a Rapid In Vitro and In Vivo Transfection System That Enables Double-Crossover Gene Knockout Studies.
C. H. M. Kocken, H. Ozwara, A. van der Wel, A. L. Beetsma, J. M. Mwenda, and A. W. Thomas (2002)
Infect. Immun. 70, 655-660
   Abstract »    Full Text »    PDF »
Topology and replication of a nuclear episomal plasmid in the rodent malaria Plasmodium berghei.
D. H. Williamson, C. J. Janse, P. W. Moore, A. P. Waters, and P. R. Preiser (2002)
Nucleic Acids Res. 30, 726-731
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Knockout of the Rodent Malaria Parasite Chitinase PbCHT1 Reduces Infectivity to Mosquitoes.
J. T. Dessens, J. Mendoza, C. Claudianos, J. M. Vinetz, E. Khater, S. Hassard, G. R. Ranawaka, and R. E. Sinden (2001)
Infect. Immun. 69, 4041-4047
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Membrane Transport in the Malaria-Infected Erythrocyte.
K. Kirk (2001)
Physiol Rev 81, 495-537
   Abstract »    Full Text »    PDF »
An alteration in concatameric structure is associated with efficient segregation of plasmids in transfected Plasmodium falciparum parasites.
R. A. O'Donnell, P. R. Preiser, D. H. Williamson, P. W. Moore, A. F. Cowman, and B. S. Crabb (2001)
Nucleic Acids Res. 29, 716-724
   Abstract »    Full Text »    PDF »
Stable Expression of a New Chimeric Fluorescent Reporter in the Human Malaria Parasite Plasmodium falciparum.
M. Kadekoppala, K. Kline, T. Akompong, and K. Haldar (2000)
Infect. Immun. 68, 2328-2332
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Targeted Disruption of the Plasmodium berghei CTRP Gene Reveals Its Essential Role in Malaria Infection of the Vector Mosquito.
M. Yuda, H. Sakaida, and Y. Chinzei (1999)
J. Exp. Med. 190, 1711-1716
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A set of independent selectable markers for transfection of the human malaria parasite Plasmodium falciparum.
C. B. Mamoun, I. Y. Gluzman, S. Goyard, S. M. Beverley, and D. E. Goldberg (1999)
PNAS 96, 8716-8720
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Transgenic Expression of a Mosquito-Stage Malarial Protein, Pbs21, in Blood Stages of Transformed Plasmodium berghei and Induction of an Immune Response upon Infection.
G. Margos, M. R. van Dijk, J. Ramesar, C. J. Janse, A. P. Waters, and R. E. Sinden (1998)
Infect. Immun. 66, 3884-3891
   Abstract »    Full Text »    PDF »
Molecular Basis of In Vivo Resistance to Sulfadoxine-Pyrimethamine in African Adult Patients Infected with Plasmodium falciparum Malaria Parasites.
L. K. Basco, R. Tahar, and P. Ringwald (1998)
Antimicrob. Agents Chemother. 42, 1811-1814
   Abstract »    Full Text »
Precise Timing of Expression of a Plasmodium falciparum-derived Transgene in Plasmodium berghei Is a Critical Determinant of Subsequent Subcellular Localization.
C. H. M. Kocken, A. M. van der Wel, M. A. Dubbeld, D. L. Narum, F. M. van de Rijke, G.-J. van Gemert, X. van der Linde, L. H. Bannister, C. Janse, A. P. Waters, et al. (1998)
J. Biol. Chem. 273, 15119-15124
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Transformation with human dihydrofolate reductase renders malaria parasites insensitive to WR99210 but does not affect the intrinsic activity of proguanil.
D. A. Fidock and T. E. Wellems (1997)
PNAS 94, 10931-10936
   Abstract »    Full Text »    PDF »
A novel alternate secretory pathway for the export of Plasmodium proteins into the host erythrocyte.
M. F. Wiser, H. N. Lanners, R. A. Bafford, and J. M. Favaloro (1997)
PNAS 94, 9108-9113
   Abstract »    Full Text »    PDF »
Transfection of the Primate Malaria Parasite Plasmodium knowlesi Using Entirely Heterologous Constructs.
A. M. v. d. Wel, A. M. Tomas, C. H.M. Kocken, P. Malhotra, C. J. Janse, A. P. Waters, and A. W. Thomas (1997)
J. Exp. Med. 185, 1499-1504
   Abstract »    Full Text »    PDF »
Antifolate-resistant mutants of Plasmodium falciparum dihydrofolate reductase.
W. Sirawaraporn, T. Sathitkul, R. Sirawaraporn, Y. Yuthavong, and D. V. Santi (1997)
PNAS 94, 1124-1129
   Abstract »    Full Text »    PDF »
Functional Equivalence of Structurally Distinct Ribosomes in the Malaria Parasite, Plasmodium berghei.
R. M. L. van Spaendonk, J. Ramesar, A. van Wigcheren, W. Eling, A. L. Beetsma, G.-J. van Gemert, J. Hooghof, C. J. Janse, and A. P. Waters (2001)
J. Biol. Chem. 276, 22638-22647
   Abstract »    Full Text »    PDF »


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