Integration of expression vectors into the ribosomal locus of Trypanosoma cruzi

The expression vectors of the protozoan parasite Trypanosoma cruzi pRIBOTEX and pTREX harbor a ribosomal promoter that improves gene expression and clone selection. Interestingly, the solely presence of this 810 bp long sequence leads to the integration of these vectors into the ribosomal locus, eve...

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Autor principal: Lorenzi, H.A
Otros Autores: Vazquez, M.P, Levin, M.J
Formato: Capítulo de libro
Lenguaje:Inglés
Publicado: Elsevier 2003
Acceso en línea:Registro en Scopus
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100 1 |a Lorenzi, H.A. 
245 1 0 |a Integration of expression vectors into the ribosomal locus of Trypanosoma cruzi 
260 |b Elsevier  |c 2003 
270 1 0 |m Levin, M.J.; Lab. Biol. Molec. Enfermedad Chagas, INGEBI - CONICET, University of Buenos Aires, Vuelta de Obligado 2490 2P, 1428 Buenos Aires, Argentina; email: mlevin@dna.uba.ar 
506 |2 openaire  |e Política editorial 
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504 |a Cano, M.I., Gruber, A., Vazquez, M., Cortes, A., Levin, M.J., Gonzalez, A., Degrave, W., Ramirez, J.L., Molecular karyotype of clone CL Brener chosen for the Trypanosoma cruzi genome project (1995) Mol. Biochem. Parasitol., 71, pp. 273-278 
504 |a Dietrich, P., Soares, M.B., Affonso, M.H., Floeter-Winter, L.M., The Trypanosoma cruzi ribosomal RNA-encoding gene: Analysis of promoter and upstream intergenic spacer sequences (1993) Gene, 125, pp. 103-107 
504 |a Fernandez, A.P., Nelson, K., Beverley, S.M., Evolution of nuclear ribosomal RNAs in kinetoplastid protozoa: Perspectives on the age and origins of parasitism (1993) Proc. Natl. Acad. Sci. USA, 90, pp. 11608-11612 
504 |a Hecker, H., Betschart, B., Burri, M., Schlimme, W., Functional morphology of trypanosome chromatin (1995) Parasitol. Today, 11, pp. 79-83 
504 |a Kelly, J., Ward, H., Miles, M., Kendall, G., A shuttle vector which facilitates the expression of transfected genes in Trypanosoma cruzi and Leishmania (1992) Nucleic Acids Res., 20, pp. 3963-3969 
504 |a Lin, Y., Keil, R., Mutations affecting RNA polymerase I-stimulated exchange and rDNA recombination in yeast (1991) Genetics, 127, pp. 31-38 
504 |a Martínez-Calvillo, S., Hernández, R., Trypanosoma cruzi ribosomal DNA: Mapping of a putative distal promoter (1994) Gene, 142, pp. 243-247 
504 |a Martínez-Calvillo, S., López, I., Hernández, R., PRIBOTEX expression vector: A pTEX derivative for a rapid selection of Trypanosoma cruzi transfectants (1997) Gene, 199, pp. 71-76 
504 |a Patnaik, P., Fang, X., Cross, G., The region encompassing the procyclic acidic repetitive protein (PARP) gene promoter plays a role in plasmid DNA replication in Trypanosoma brucei (1994) Nucleic Acids Res., 22, pp. 4111-4118 
504 |a Porto, R.M., Rogério Amino, R., Quartim Elias, M.C., Faria, M., Schenkman, S., Histone H1 is phosphorylated in non-replicating and infective forms of Trypanosoma cruzi (2002) Mol. Biochem. Parasitol., 119, pp. 265-271 
504 |a Stewart, S.E., Roeder, G., Transcription by RNA polymerase I stimulates mitotic recombination in Saccharomyces cerevisiae (1989) Mol. Cell. Biol., 8, pp. 3464-3472 
504 |a Tyler-Cross, R., Short, S., Floeter-Winter, L., Buck, G., Transient expression mediated by the Trypanosoma cruzi rRNA promoter (1995) Mol. Biochem. Parasitol., 72, pp. 23-31 
504 |a Vanhamme, L., Pays, E., Control of gene expression in trypanosomes (1995) Microbiol. Rev., 59, pp. 223-240 
504 |a Vázquez, M., Levin, M.J., Functional analysis of the intergenic regions of TcP2b gene loci allowed the construction of an improved Trypanosoma cruzi expression vector (1999) Gene, 239, pp. 217-225 
504 |a Voelkel-Meiman, K., Keil, R., Roeder, G., Recombination-stimulating sequences in yeast ribosomal DNA correspond to sequences regulating transcription by RNA polymerase I (1987) Cell, 48, pp. 1071-1079 
504 |a Wen, L.M., Xu, P., Benegal, G., Carvaho, M.R., Butler, D.R., Buck, G.A., Trypanosoma cruzi: Exogenously regulated gene expression (2001) Exp. Parasitol., 97, pp. 196-204 
520 3 |a The expression vectors of the protozoan parasite Trypanosoma cruzi pRIBOTEX and pTREX harbor a ribosomal promoter that improves gene expression and clone selection. Interestingly, the solely presence of this 810 bp long sequence leads to the integration of these vectors into the ribosomal locus, even though circular plasmids are poorly recombinogenic. Initially, it was suggested that a 174 bp long ribosomal-specific repeat element present in the ribosomal promoter region could be responsible for the genetic exchange. On the contrary, we demonstrate that recombination of pTREX occurs within a 86 bp long region located 120 bp downstream the transcription start point (tsp1) of the ribosomal promoter, and it does not depend on the presence of the ribosomal repeat. We also determined that a 291 bp segment encompassing the tsp1 and the 86 bp long recombination region contains all necessary signals to drive transcription and complete recombination into the rRNA locus. Finally, we demonstrate that the integration of pTREX derived plasmids into the nuclear genome occurs within the first 5 h post-transfection, and that non-integrated copies are rapidly degraded. © 2003 Elsevier Science B.V. All rights reserved.  |l eng 
536 |a Detalles de la financiación: Universidad de Buenos Aires, 01/IX-17 
536 |a Detalles de la financiación: Howard Hughes Medical Institute 
536 |a Detalles de la financiación: Fondo para la Investigación Científica y Tecnológica, PICT 01-06803 
536 |a Detalles de la financiación: Consejo Nacional de Investigaciones Científicas y Técnicas 
536 |a Detalles de la financiación: We would like to thank Maria de los Angeles Curto and graduate student Ignacio Schor for their technical assistance. This work was supported by grants from T. cruzi Genome Project of the University of Buenos Aires; Project 01/IX-17; UNDP/World Bank/WHO Special Programme for Research and Training in Tropical Diseases; Proyecto Genoma de T. cruzi of Centro Argentino-Brasileño de Biotecnologı́a (CABBIO), CONICET, and FONCYT project PICT 01-06803. In addition, the work of MJL was partially supported by an International Research Scholar grant from the Howard Hughes Medical Institute, Chevy Chase, MD, USA. 
593 |a Lab. Biol. Molec. Enfermedad Chagas, INGEBI - CONICET, University of Buenos Aires, Vuelta de Obligado 2490 2P, 1428 Buenos Aires, Argentina 
690 1 0 |a EXPRESSION VECTOR 
690 1 0 |a RECOMBINATION 
690 1 0 |a RIBOSOMAL PROMOTER 
690 1 0 |a TRANSFECTION 
690 1 0 |a PLASMID VECTOR 
690 1 0 |a ARTICLE 
690 1 0 |a EXPRESSION VECTOR 
690 1 0 |a GENE LOCATION 
690 1 0 |a GENE LOCUS 
690 1 0 |a GENETIC RECOMBINATION 
690 1 0 |a GENETIC TRANSFECTION 
690 1 0 |a GENOME 
690 1 0 |a NONHUMAN 
690 1 0 |a NUCLEOTIDE REPEAT 
690 1 0 |a NUCLEOTIDE SEQUENCE 
690 1 0 |a PRIORITY JOURNAL 
690 1 0 |a PROMOTER REGION 
690 1 0 |a RIBOSOME 
690 1 0 |a RNA RECOMBINATION 
690 1 0 |a TIME 
690 1 0 |a TRANSCRIPTION INITIATION SITE 
690 1 0 |a TRANSCRIPTION REGULATION 
690 1 0 |a TRYPANOSOMA CRUZI 
690 1 0 |a PROTOZOA 
690 1 0 |a TRYPANOSOMA 
690 1 0 |a TRYPANOSOMA CRUZI 
690 1 0 |a TRYPANOSOMA CRUZI 
700 1 |a Vazquez, M.P. 
700 1 |a Levin, M.J. 
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