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Science 26 May 1995:
Vol. 268. no. 5214, pp. 1179 - 1183
DOI: 10.1126/science.7761835
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Articles

Science, Vol 268, Issue 5214, 1179-1183
Copyright © 1995 by American Association for the Advancement of Science

articles

Inducible gene expression in trypanosomes mediated by a prokaryotic repressor

E Wirtz and C Clayton

Zentrum fur Molekulare Biologie, Universitat Heidelberg, Germany.

An inducible expression system was developed for the protozoan parasite Trypanosoma brucei. Transgenic trypanosomes expressing the tetracycline repressor of Escherichia coli exhibited inducer (tetracycline)-dependent expression of chromosomally integrated reporter genes under the control of a procyclic acidic repetitive protein (PARP) promoter bearing a tet operator. Reporter expression could be controlled over a range of four orders of magnitude in response to tetracycline concentration, a degree of regulation that exceeds those exhibited by other eukaryotic repression-based systems. The tet repressor-controlled PARP promoter should be a valuable tool for the study of trypanosome biochemistry, pathogenicity, and cell and molecular biology.


THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
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B. G. Oliver, P. M. Silver, C. Marie, S. J. Hoot, S. E. Leyde, and T. C. White (2008)
Microbiology 154, 960-970
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Intraflagellar Transport and Functional Analysis of Genes Required for Flagellum Formation in Trypanosomes.
S. Absalon, T. Blisnick, L. Kohl, G. Toutirais, G. Dore, D. Julkowska, A. Tavenet, and P. Bastin (2008)
Mol. Biol. Cell 19, 929-944
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Sphingolipid synthesis is necessary for kinetoplast segregation and cytokinesis in Trypanosoma brucei.
A. Fridberg, C. L. Olson, E. S. Nakayasu, K. M. Tyler, I. C. Almeida, and D. M. Engman (2008)
J. Cell Sci. 121, 522-535
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Adenosine Kinase of Trypanosoma brucei and Its Role in Susceptibility to Adenosine Antimetabolites.
A. Luscher, P. Onal, A.-M. Schweingruber, and P. Maser (2007)
Antimicrob. Agents Chemother. 51, 3895-3901
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Depletion of newly synthesized Argonaute1 impairs the RNAi response in Trypanosoma brucei.
H. Shi, C. Tschudi, and E. Ullu (2007)
RNA 13, 1132-1139
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An Essential Cell Cycle-regulated Nucleolar Protein Relocates to the Mitotic Spindle Where It Is Involved in Mitotic Progression in Trypanosoma brucei.
N. Boucher, D. Dacheux, C. Giroud, and T. Baltz (2007)
J. Biol. Chem. 282, 13780-13790
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Mitochondrial Fatty Acid Synthesis in Trypanosoma brucei.
J. L. Stephens, S. H. Lee, K. S. Paul, and P. T. Englund (2007)
J. Biol. Chem. 282, 4427-4436
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Knock-downs of Iron-Sulfur Cluster Assembly Proteins IscS and IscU Down-regulate the Active Mitochondrion of Procyclic Trypanosoma brucei.
O. Smid, E. Horakova, V. Vilimova, I. Hrdy, R. Cammack, A. Horvath, J. Lukes, and J. Tachezy (2006)
J. Biol. Chem. 281, 28679-28686
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Molecular Characterization of Trypanosoma brucei P-type H+-ATPases.
S. Luo, J. Fang, and R. Docampo (2006)
J. Biol. Chem. 281, 21963-21973
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Overexpression of a Cytochrome b5 Reductase-like Protein Causes Kinetoplast DNA Loss in Trypanosoma brucei.
S. A. Motyka, M. E. Drew, G. Yildirir, and P. T. Englund (2006)
J. Biol. Chem. 281, 18499-18506
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Experimental and in Silico Analyses of Glycolytic Flux Control in Bloodstream Form Trypanosoma brucei.
M.-A. Albert, J. R. Haanstra, V. Hannaert, J. Van Roy, F. R. Opperdoes, B. M. Bakker, and P. A. M. Michels (2005)
J. Biol. Chem. 280, 28306-28315
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Mitochondrial Initiation Factor 2 of Trypanosoma brucei Binds Imported Formylated Elongator-type tRNAMet.
F. Charriere, T. H. P. Tan, and A. Schneider (2005)
J. Biol. Chem. 280, 15659-15665
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Tetracycline analogue-regulated transgene expression in Plasmodium falciparum blood stages using Toxoplasma gondii transactivators.
M. Meissner, E. Krejany, P. R. Gilson, T. F. de Koning-Ward, D. Soldati, and B. S. Crabb (2005)
PNAS 102, 2980-2985
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Regulated gene expression by glucocorticoids in cultured Virginia pine (Pinus virginiana Mill.) cells.
W. Tang and R. J. Newton (2004)
J. Exp. Bot. 55, 1499-1508
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Protein Targeting of an Unusual, Evolutionarily Conserved Adenylate Kinase to a Eukaryotic Flagellum.
T. J. Pullen, M. L. Ginger, S. J. Gaskell, and K. Gull (2004)
Mol. Biol. Cell 15, 3257-3265
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The Small Chromosomes of Trypanosoma brucei Involved in Antigenic Variation Are Constructed Around Repetitive Palindromes.
B. Wickstead, K. Ersfeld, and K. Gull (2004)
Genome Res. 14, 1014-1024
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Trypanosoma brucei Plasma Membrane-Type Ca2+-ATPase 1 (TbPMC1) and 2 (TbPMC2) Genes Encode Functional Ca2+-ATPases Localized to the Acidocalcisomes and Plasma Membrane, and Essential for Ca2+ Homeostasis and Growth.
S. Luo, P. Rohloff, J. Cox, S. A. Uyemura, and R. Docampo (2004)
J. Biol. Chem. 279, 14427-14439
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The frequency of gene targeting in Trypanosoma brucei is independent of target site copy number.
B. Wickstead, K. Ersfeld, and K. Gull (2003)
Nucleic Acids Res. 31, 3993-4000
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Ex Vivo and In Vitro Identification of a Consensus Promoter for VSG Genes Expressed by Metacyclic-Stage Trypanosomes in the Tsetse Fly.
M. L. Ginger, P. A. Blundell, A. M. Lewis, A. Browitt, A. Gunzl, and J. D. Barry (2002)
Eukaryot. Cell 1, 1000-1009
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Expression of the human DNA glycosylase hSMUG1 in Trypanosoma brucei causes DNA damage and interferes with J biosynthesis.
S. Ulbert, M. Cross, R. J. Boorstein, G. W. Teebor, and P. Borst (2002)
Nucleic Acids Res. 30, 3919-3926
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Mitochondrial Substrate Level Phosphorylation Is Essential for Growth of Procyclic Trypanosoma brucei.
N. Bochud-Allemann and A. Schneider (2002)
J. Biol. Chem. 277, 32849-32854
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Trypanin is a cytoskeletal linker protein and is required for cell motility in African trypanosomes.
N. R. Hutchings, J. E. Donelson, and K. L. Hill (2002)
J. Cell Biol. 156, 867-877
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A novel GFP approach for the analysis of genetic exchange in trypanosomes allowing the in situ detection of mating events.
L. E. H. Bingle, J. L. Eastlake, M. Bailey, and W. C. Gibson (2001)
Microbiology 147, 3231-3240
   Abstract »    Full Text »    PDF »
Degradation of the unstable EP1 mRNA in Trypanosoma brucei involves initial destruction of the 3'-untranslated region.
H. Irmer and C. Clayton (2001)
Nucleic Acids Res. 29, 4707-4715
   Abstract »    Full Text »    PDF »
Modulation of myosin A expression by a newly established tetracycline repressor-based inducible system in Toxoplasma gondii.
M. Meissner, S. Brecht, H. Bujard, and D. Soldati (2001)
Nucleic Acids Res. 29, e115
   Abstract »    Full Text »    PDF »
Targeting of cytoskeletal proteins to the flagellum of Trypanosoma brucei.
K Ersfeld and K Gull (2001)
J. Cell Sci. 114, 141-148
   Abstract »    PDF »
Isolation of Trypanosoma brucei CYC2 and CYC3 Cyclin Genes by Rescue of a Yeast G1 Cyclin Mutant. FUNCTIONAL CHARACTERIZATION OF CYC2.
J. J. Van Hellemond, P. Neuville, R. T. Schwarz, K. R. Matthews, and J. C. Mottram (2000)
J. Biol. Chem. 275, 8315-8323
   Abstract »    Full Text »    PDF »
Flagellum ontogeny in trypanosomes studied via an inherited and regulated RNA interference system.
P Bastin, K Ellis, L Kohl, and K Gull (2000)
J. Cell Sci. 113, 3321-3328
   Abstract »    PDF »
Flagellar Morphogenesis: Protein Targeting and Assembly in the Paraflagellar Rod of Trypanosomes.
P. Bastin, T. H. MacRae, S. B. Francis, K. R. Matthews, and K. Gull (1999)
Mol. Cell. Biol. 19, 8191-8200
   Abstract »    Full Text »    PDF »
Double-stranded RNA induces mRNA degradation in Trypanosoma brucei.
H. Ngo, C. Tschudi, K. Gull, and E. Ullu (1998)
PNAS 95, 14687-14692
   Abstract »    Full Text »    PDF »
Activity of a Trypanosome Metacyclic Variant Surface Glycoprotein Gene Promoter Is Dependent upon Life Cycle Stage and Chromosomal Context.
S. V. Graham, B. Wymer, and J. D. Barry (1998)
Mol. Cell. Biol. 18, 1137-1146
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Inhibition of Trypanosoma brucei Gene Expression by RNA Interference Using an Integratable Vector with Opposing T7 Promoters.
Z. Wang, J. C. Morris, M. E. Drew, and P. T. Englund (2000)
J. Biol. Chem. 275, 40174-40179
   Abstract »    Full Text »    PDF »
Trypanin is a cytoskeletal linker protein and is required for cell motility in African trypanosomes.
N. R. Hutchings, J. E. Donelson, and K. L. Hill (2002)
J. Cell Biol. 156, 867-877
   Abstract »    Full Text »    PDF »


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