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Abstract:

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.

Registro:

Documento: Artículo
Título:Integration of expression vectors into the ribosomal locus of Trypanosoma cruzi
Autor:Lorenzi, H.A.; Vazquez, M.P.; Levin, M.J.
Filiación:Lab. Biol. Molec. Enfermedad Chagas, INGEBI - CONICET, University of Buenos Aires, Vuelta de Obligado 2490 2P, 1428 Buenos Aires, Argentina
Palabras clave:Expression vector; Recombination; Ribosomal promoter; Transfection; plasmid vector; article; expression vector; gene location; gene locus; genetic recombination; genetic transfection; genome; nonhuman; nucleotide repeat; nucleotide sequence; priority journal; promoter region; ribosome; RNA recombination; time; transcription initiation site; transcription regulation; Trypanosoma cruzi; Protozoa; Trypanosoma; Trypanosoma cruzi; Trypanosoma cruzi
Año:2003
Volumen:310
Número:1-2
Página de inicio:91
Página de fin:99
DOI: http://dx.doi.org/10.1016/S0378-1119(03)00502-X
Título revista:Gene
Título revista abreviado:Gene
ISSN:03781119
CODEN:GENED
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03781119_v310_n1-2_p91_Lorenzi

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Citas:

---------- APA ----------
Lorenzi, H.A., Vazquez, M.P. & Levin, M.J. (2003) . Integration of expression vectors into the ribosomal locus of Trypanosoma cruzi. Gene, 310(1-2), 91-99.
http://dx.doi.org/10.1016/S0378-1119(03)00502-X
---------- CHICAGO ----------
Lorenzi, H.A., Vazquez, M.P., Levin, M.J. "Integration of expression vectors into the ribosomal locus of Trypanosoma cruzi" . Gene 310, no. 1-2 (2003) : 91-99.
http://dx.doi.org/10.1016/S0378-1119(03)00502-X
---------- MLA ----------
Lorenzi, H.A., Vazquez, M.P., Levin, M.J. "Integration of expression vectors into the ribosomal locus of Trypanosoma cruzi" . Gene, vol. 310, no. 1-2, 2003, pp. 91-99.
http://dx.doi.org/10.1016/S0378-1119(03)00502-X
---------- VANCOUVER ----------
Lorenzi, H.A., Vazquez, M.P., Levin, M.J. Integration of expression vectors into the ribosomal locus of Trypanosoma cruzi. Gene. 2003;310(1-2):91-99.
http://dx.doi.org/10.1016/S0378-1119(03)00502-X