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Science 5 November 1993:
Vol. 262. no. 5135, pp. 911 - 914
DOI: 10.1126/science.8235614
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Articles

Science, Vol 262, Issue 5135, 911-914
Copyright © 1993 by American Association for the Advancement of Science

articles

Gene replacement in Toxoplasma gondii with chloramphenicol acetyltransferase as selectable marker

K Kim, D Soldati, and JC Boothroyd

Department of Microbiology and Immunology, Stanford University School of Medicine 94305.

A system for stable transformation of Toxoplasma gondii tachyzoites was developed that exploited the susceptibility of Toxoplasma to chloramphenicol. Introduction of the chloramphenicol acetyltransferase (CAT) gene fused to Toxoplasma flanking sequences followed by chloramphenicol selection resulted in parasites stably expressing CAT. A construct incorporating the tandemly repeated gene, B1, targeted efficiently to its homologous chromosomal locus. Knockout of the single-copy gene, ROP1, was also successful. Stable transformation should permit the identification and analysis of Toxoplasma genes important in the interaction of this opportunistic parasite with its host.


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Microneme protein 8 - a new essential invasion factor in Toxoplasma gondii.
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Differential Localization of Alternatively Spliced Hypoxanthine-Xanthine-Guanine Phosphoribosyltransferase Isoforms in Toxoplasma gondii.
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J. Cell Biol. 165, 653-662
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Identification of the membrane receptor of a class XIV myosin in Toxoplasma gondii.
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J. Cell Biol. 165, 383-393
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Comparison of Two Widely Used PCR Primer Systems for Detection of Toxoplasma in Amniotic Fluid, Blood, and Tissues.
E. Chabbert, L. Lachaud, L. Crobu, and P. Bastien (2004)
J. Clin. Microbiol. 42, 1719-1722
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Pleiotropic effect due to targeted depletion of secretory rhoptry protein ROP2 in Toxoplasma gondii.
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High-Throughput Growth Assay for Toxoplasma gondii Using Yellow Fluorescent Protein.
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Biogenesis of Nanotubular Network in Toxoplasma Parasitophorous Vacuole Induced by Parasite Proteins.
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Toxoplasma gondii Cyclic GMP-Dependent Kinase: Chemotherapeutic Targeting of an Essential Parasite Protein Kinase.
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Modulation of myosin A expression by a newly established tetracycline repressor-based inducible system in Toxoplasma gondii.
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Identification and Characterization of an Escorter for Two Secretory Adhesins in Toxoplasma gondii.
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Expression of Herpes Simplex Virus Thymidine Kinase in Toxoplasma gondii Attenuates Tachyzoite Virulence in Mice.
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Transformed Toxoplasma gondii Tachyzoites Expressing the Circumsporozoite Protein of Plasmodium knowlesi Elicit a Specific Immune Response in Rhesus Monkeys.
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Transmembrane Insertion of the Toxoplasma gondii GRA5 Protein Occurs after Soluble Secretion into the Host Cell.
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Targeted Reduction of Nucleoside Triphosphate Hydrolase by Antisense RNA Inhibits Toxoplasma gondii Proliferation.
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J. Biol. Chem. 274, 5083-5087
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Targeted Disruption of the GRA2 Locus in Toxoplasma gondii Decreases Acute Virulence in Mice.
C. Mercier, D. K. Howe, D. Mordue, M. Lingnau, and L. D. Sibley (1998)
Infect. Immun. 66, 4176-4182
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The parasitophorous vacuole membrane surrounding Plasmodium and Toxoplasma: an unusual compartment in infected cells.
K Lingelbach and K. Joiner (1998)
J. Cell Sci. 111, 1467-1475
   Abstract »    PDF »
Analysis of Toxoplasma gondii stably transfected with a transmembrane variant of its major surface protein, SAG1.
F Seeber, J. Dubremetz, and J. Boothroyd (1998)
J. Cell Sci. 111, 23-29
   Abstract »    PDF »
Insertional Tagging, Cloning, and Expression of the Toxoplasma gondii Hypoxanthine-Xanthine-Guanine Phosphoribosyltransferase Gene. USE AS A SELECTABLE MARKER FOR STABLE TRANSFORMATION.
R. G.K. Donald, D. Carter, B. Ullman, and D. S. Roos (1996)
J. Biol. Chem. 271, 14010-14019
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Stable transfection of malaria parasite blood stages.
M. van Dijk, A. Waters, and C. Janse (1995)
Science 268, 1358-1362
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Unusual strategies of gene expression and control in parasites.
T. Nilsen (1994)
Science 264, 1868-1869
   PDF »
Daughter Cell Assembly in the Protozoan Parasite Toxoplasma gondii.
K. Hu, T. Mann, B. Striepen, C. J. M. Beckers, D. S. Roos, and J. M. Murray (2002)
Mol. Biol. Cell 13, 593-606
   Abstract »    Full Text »    PDF »


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