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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| Acceso en línea: | http://hdl.handle.net/20.500.12110/paper_03781119_v310_n1-2_p91_Lorenzi |
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todo:paper_03781119_v310_n1-2_p91_Lorenzi2023-10-03T15:31:48Z Integration of expression vectors into the ribosomal locus of Trypanosoma cruzi Lorenzi, H.A. Vazquez, M.P. Levin, M.J. 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 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. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_03781119_v310_n1-2_p91_Lorenzi |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
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 |
| spellingShingle |
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 Lorenzi, H.A. Vazquez, M.P. Levin, M.J. Integration of expression vectors into the ribosomal locus of Trypanosoma cruzi |
| topic_facet |
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 |
| description |
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. |
| format |
JOUR |
| author |
Lorenzi, H.A. Vazquez, M.P. Levin, M.J. |
| author_facet |
Lorenzi, H.A. Vazquez, M.P. Levin, M.J. |
| author_sort |
Lorenzi, H.A. |
| title |
Integration of expression vectors into the ribosomal locus of Trypanosoma cruzi |
| title_short |
Integration of expression vectors into the ribosomal locus of Trypanosoma cruzi |
| title_full |
Integration of expression vectors into the ribosomal locus of Trypanosoma cruzi |
| title_fullStr |
Integration of expression vectors into the ribosomal locus of Trypanosoma cruzi |
| title_full_unstemmed |
Integration of expression vectors into the ribosomal locus of Trypanosoma cruzi |
| title_sort |
integration of expression vectors into the ribosomal locus of trypanosoma cruzi |
| url |
http://hdl.handle.net/20.500.12110/paper_03781119_v310_n1-2_p91_Lorenzi |
| work_keys_str_mv |
AT lorenziha integrationofexpressionvectorsintotheribosomallocusoftrypanosomacruzi AT vazquezmp integrationofexpressionvectorsintotheribosomallocusoftrypanosomacruzi AT levinmj integrationofexpressionvectorsintotheribosomallocusoftrypanosomacruzi |
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