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Science 14 December 1990:
Vol. 250. no. 4987, pp. 1583 - 1587
DOI: 10.1126/science.2177225
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Articles

Science, Vol 250, Issue 4987, 1583-1587
Copyright © 1990 by American Association for the Advancement of Science

articles

Homologous recombination and stable transfection in the parasitic protozoan Trypanosoma brucei

MG Lee and LH Van der Ploeg

Division of Tropical Medicine, School of Public Health, Columbia University, New York, NY 10032.

Development of methods for the manipulation of the genomes of parasitic protozoa will lead to enhanced understanding of parasite biology and host-parasite relationships. Efficient gene transfer and targeted integration by homologous recombination were achieved in the parasitic protozoan Trypanosoma brucei, the causative agent of sleeping sickness. An expression vector with the neomycin phosphotransferase gene (neo), under the control of a procyclic acidic repetitive protein (PARP) gene promoter, was targeted into an intergenic region in beta alpha-tubulin-gene tandem array. Sixteen copies of neo were found in a tandem array in one of the transfectants where the PARP promoter controlled alpha-amanitin-resistant transcription of neo, whereas transcription of tubulin genes remained alpha-amanitin-sensitive.


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The frequency of gene targeting in Trypanosoma brucei is independent of target site copy number.
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The Two RNA Ligases of the Trypanosoma brucei RNA Editing Complex: Cloning the Essential Band IV Gene and Identifying the Band V Gene.
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Determinants for cap trimethylation of the U2 small nuclear RNA are not conserved between Trypanosoma brucei and higher eukaryotic organisms.
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Flagellar Morphogenesis: Protein Targeting and Assembly in the Paraflagellar Rod of Trypanosomes.
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A role for RAD51 and homologous recombination in Trypanosoma brucei antigenic variation.
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In Vitro Analysis of alpha -Amanitin-Resistant Transcription from the rRNA, Procyclic Acidic Repetitive Protein, and Variant Surface Glycoprotein Gene Promoters in Trypanosoma brucei.
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Protein transport and flagellum assembly dynamics revealed by analysis of the paralysed trypanosome mutant snl-1.
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The GPI-Phospholipase C of Trypanosoma brucei Is Nonessential But Influences Parasitemia in Mice.
H. Webb, N. Carnall, L. Vanhamme, S. Rolin, J. V. D. Abbeele, S. Welburn, E. Pays, and M. Carrington (1997)
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Creation of Homozygous Mutants of Leishmania donovani with Single Targeting Constructs.
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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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Inducible gene expression in trypanosomes mediated by a prokaryotic repressor.
E Wirtz and C Clayton (1995)
Science 268, 1179-1183
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Transient transfection and expression in the obligate intracellular parasite Toxoplasma gondii.
D Soldati and J. Boothroyd (1993)
Science 260, 349-352
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Control of Antigenic Variation in African Trypanosomas.
P. Borst, J.H. Gommers-Ampt, M.J.L. Ligtenberg, G. Rudenko, R. Kieft, M.C. Taylor, P.A. Blundell, and F. van Leeuwen (1993)
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Homologous recombination in the tandem calmodulin genes of Trypanosoma brucei yields multiple products: compensation for deleterious deletions by gene amplification..
J E Eid and B Sollner-Webb (1991)
Genes & Dev. 5, 2024-2032
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