Nucleic Acids Research, 1992, Vol. 20, No. 15 3963-3969
© 1992
Methods |
A shuttle vector which facilitates the expression of transfected genes in Trypanosoma cruzi and Leishmania
Department of Medical parasitology, London school of Hygiene and Tropical Medicine Keppel Street, London WC1E 7HT, UK
Received May 5, 1992. Revised July 6, 1992. Accepted July 6, 1992.
A Trypanosoma cruzi expression vector has been constructed using sequences derived from the flanking regions of the glyceraldehyde 3-phosphate dehydrogenase (gGAPDH) genes. The neomycln phosphotransferase (neor) gene was incorporated as a selectable marker. Using electroporatlon we have introduced this vector into both T.cruzl and Leishmania cells and conferred G418 resistance. Transformation is mediated by large extrachromosomal circular elements composed of head-to-tail tandem repeats of the vector. The transformed phenotype is stable for at least 6 months in the absence of G418 and can be maintained during passage through the T.cruzl ifecycle. Foreign genes Inserted into an expression site within the vector (pTEX) can be expressed at high levels In transformed cells. To our knowledge this paper describes the first trypanosome shuttle vector and the first vector which functions in both trypanosomes and Leishmania.
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  L. L Penha, C. B Sant'Anna, L. Mendonca-Previato, N. L Cunha-e-Silva, J. O Previato, and A. P. C A Lima Sorting of phosphoglucomutase to glycosomes in Trypanosoma cruzi is mediated by an internal domain Glycobiology, December 1, 2009; 19(12): 1462 - 1472. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Dallagiovanna, A. Correa, C. M. Probst, F. Holetz, P. Smircich, A. M. de Aguiar, F. Mansur, C. V. da Silva, R. A. Mortara, B. Garat, et al. Functional Genomic Characterization of mRNAs Associated with TcPUF6, a Pumilio-like Protein from Trypanosoma cruzi J. Biol. Chem., March 28, 2008; 283(13): 8266 - 8273. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. R. Heras, M. C. Lopez, M. Olivares, and M. C. Thomas The L1Tc non-LTR retrotransposon of Trypanosoma cruzi contains an internal RNA-pol II-dependent promoter that strongly activates gene transcription and generates unspliced transcripts Nucleic Acids Res., April 1, 2007; 35(7): 2199 - 2214. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Chamekh, V. Vercruysse, M. Habib, M. Lorent, M. Goldman, A. Allaoui, and B. Vray Transfection of Trypanosoma cruzi with Host CD40 Ligand Results in Improved Control of Parasite Infection Infect. Immun., October 1, 2005; 73(10): 6552 - 6561. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Okura, J. Fang, M. L. Salto, R. S. Singer, R. Docampo, and S. N. J. Moreno A Lipid-modified Phosphoinositide-specific Phospholipase C (TcPI-PLC) Is Involved in Differentiation of Trypomastigotes to Amastigotes of Trypanosoma cruzi J. Biol. Chem., April 22, 2005; 280(16): 16235 - 16243. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. C. Santos, C. Sant'Anna, A. Terres, N. L. Cunha-e-Silva, J. Scharfstein, and A. P. C. de A. Lima Chagasin, the endogenous cysteine-protease inhibitor of Trypanosoma cruzi, modulates parasite differentiation and invasion of mammalian cells J. Cell Sci., March 1, 2005; 118(5): 901 - 915. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. O. Obado, M. C. Taylor, S. R. Wilkinson, E. V. Bromley, and J. M. Kelly Functional mapping of a trypanosome centromere by chromosome fragmentation identifies a 16-kb GC-rich transcriptional "strand-switch" domain as a major feature Genome Res., January 1, 2005; 15(1): 36 - 43. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Bhatia, M. Sinha, B. Luxon, and N. Garg Utility of the Trypanosoma cruzi Sequence Database for Identification of Potential Vaccine Candidates by In Silico and In Vitro Screening Infect. Immun., November 1, 2004; 72(11): 6245 - 6254. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. M. Manque, I. Neira, V. D. Atayde, E. Cordero, A. T. Ferreira, J. F. da Silveira, M. Ramirez, and N. Yoshida Cell Adhesion and Ca2+ Signaling Activity in Stably Transfected Trypanosoma cruzi Epimastigotes Expressing the Metacyclic Stage-Specific Surface Molecule gp82 Infect. Immun., March 1, 2003; 71(3): 1561 - 1565. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. P. Price, M. R. Menon, C. Panethymitaki, D. Goulding, P. G. McKean, and D. F. Smith Myristoyl-CoA:Protein N-Myristoyltransferase, an Essential Enzyme and Potential Drug Target in Kinetoplastid Parasites J. Biol. Chem., February 21, 2003; 278(9): 7206 - 7214. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. R. Wilkinson, S. O. Obado, I. L. Mauricio, and J. M. Kelly Trypanosoma cruzi expresses a plant-like ascorbate-dependent hemoperoxidase localized to the endoplasmic reticulum PNAS, October 15, 2002; 99(21): 13453 - 13458. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. R. Wilkinson, D. J. Meyer, M. C. Taylor, E. V. Bromley, M. A. Miles, and J. M. Kelly The Trypanosoma cruzi Enzyme TcGPXI Is a Glycosomal Peroxidase and Can Be Linked to Trypanothione Reduction by Glutathione or Tryparedoxin J. Biol. Chem., May 3, 2002; 277(19): 17062 - 17071. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. H. Williamson, C. J. Janse, P. W. Moore, A. P. Waters, and P. R. Preiser Topology and replication of a nuclear episomal plasmid in the rodent malaria Plasmodium berghei Nucleic Acids Res., February 1, 2002; 30(3): 726 - 731. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. A. O'Donnell, P. R. Preiser, D. H. Williamson, P. W. Moore, A. F. Cowman, and B. S. Crabb An alteration in concatameric structure is associated with efficient segregation of plasmids in transfected Plasmodium falciparum parasites Nucleic Acids Res., February 1, 2001; 29(3): 716 - 724. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Büchner, P. Roth, G. Schotta, V. Krauss, H. Saumweber, G. Reuter, and R. Dorn Genetic and Molecular Complexity of the Position Effect Variegation Modifier mod(mdg4) in Drosophila Genetics, May 1, 2000; 155(1): 141 - 157. [Abstract] [Full Text]
|
|
|
|
|
|
J. M. Di Noia, I. D'Orso, D. O. Sanchez, and A. C. C. Frasch AU-rich Elements in the 3'-Untranslated Region of a New Mucin-type Gene Family of Trypanosoma cruzi Confers mRNA Instability and Modulates Translation Efficiency J. Biol. Chem., March 31, 2000; 275(14): 10218 - 10227. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Bakalara, X. Santarelli, C. Davis, and T. Baltz Purification, Cloning, and Characterization of an Acidic Ectoprotein Phosphatase Differentially Expressed in the Infectious Bloodstream Form of Trypanosoma brucei J. Biol. Chem., March 17, 2000; 275(12): 8863 - 8871. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. R. Wilkinson, N. J. Temperton, A. Mondragon, and J. M. Kelly Distinct Mitochondrial and Cytosolic Enzymes Mediate Trypanothione-dependent Peroxide Metabolism in Trypanosoma cruzi J. Biol. Chem., March 10, 2000; 275(11): 8220 - 8225. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Brooks, L Tetley, G. Coombs, and J. Mottram Processing and trafficking of cysteine proteases in Leishmania mexicana J. Cell Sci., January 11, 2000; 113(22): 4035 - 4041. [Abstract] [PDF]
|
|
|
|
|
|
J. A. Huete-Perez, J. C. Engel, L. S. Brinen, J. C. Mottram, and J. H. McKerrow Protease Trafficking in Two Primitive Eukaryotes Is Mediated by a Prodomain Protein Motif J. Biol. Chem., June 4, 1999; 274(23): 16249 - 16256. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y. Miyahira, S. Kobayashi, T. Takeuchi, T. Kamiyama, T. Nara, J. Nakajima-Shimada, and T. Aoki Induction of CD8+ T cell-mediated protective immunity against Trypanosoma cruzi Int. Immunol., February 1, 1999; 11(2): 133 - 141. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Maga, T Sherwin, S Francis, K Gull, and J. LeBowitz Genetic dissection of the Leishmania paraflagellar rod, a unique flagellar cytoskeleton structure J. Cell Sci., January 8, 1999; 112(16): 2753 - 2763. [Abstract] [PDF]
|
|
|
|
|
|
K. A. Norris Stable Transfection of Trypanosoma cruzi Epimastigotes with the Trypomastigote-Specific Complement Regulatory Protein cDNA Confers Complement Resistance Infect. Immun., June 1, 1998; 66(6): 2460 - 2465. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. W. Black and J. C. Boothroyd Development of a Stable Episomal Shuttle Vector for Toxoplasma gondii J. Biol. Chem., February 13, 1998; 273(7): 3972 - 3979. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. C. Mottram, M. J. Frame, D. R. Brooks, L. Tetley, J. E. Hutchison, A. E. Souza, and G. H. Coombs The Multiple cpb Cysteine Proteinase Genes of Leishmania mexicana Encode Isoenzymes That Differ in Their Stage Regulation and Substrate Preferences J. Biol. Chem., May 30, 1997; 272(22): 14285 - 14293. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Garg, R. L. Tarleton, and K. Mensa-Wilmot Proteins with Glycosylphosphatidylinositol (GPI) Signal Sequences Have Divergent Fates during a GPI Deficiency. GPIs ARE ESSENTIAL FOR NUCLEAR DIVISION IN TRYPANOSOMA CRUZI J. Biol. Chem., May 9, 1997; 272(19): 12482 - 12491. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. M. R. Teixeira, L. V. Kirchhoff, and J. E. Donelson Post-transcriptional Elements Regulating Expression of mRNAs from the Amastin/Tuzin Gene Cluster of Trypanosoma cruzi J. Biol. Chem., September 22, 1995; 270(38): 22586 - 22594. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
I. D'Orso and A. C. C. Frasch Functionally Different AU- and G-rich cis-Elements Confer Developmentally Regulated mRNA Stability in Trypanosoma cruzi by Interaction with Specific RNA-binding Proteins J. Biol. Chem., May 4, 2001; 276(19): 15783 - 15793. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Furuya, M. Okura, F. A. Ruiz, D. A. Scott, and R. Docampo TcSCA Complements Yeast Mutants Defective in Ca2+ Pumps and Encodes a Ca2+-ATPase That Localizes to the Endoplasmic Reticulum of Trypanosoma cruzi J. Biol. Chem., August 24, 2001; 276(35): 32437 - 32445. [Abstract] [Full Text] [PDF]
|
|