Artículo

DaRocha, W.D.; Silva, R.A.; Bartholomeu, D.C.; Pires, S.F.; Freitas, J.M.; Macedo, A.M.; Vazquez, M.P.; Levin, M.J.; Teixeira, S.M.R. "Expression of exogenous genes in Trypanosoma cruzi: Improving vectors and electroporation protocols" (2004) Parasitology Research. 92(2):113-120
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Abstract:

To improve transfection efficiency in Trypanosoma cruzi, we developed a new electroporation protocol and expression vectors which use luciferase and green and red fluorescent proteins as reporter genes. In transient transfections, the electroporation conditions reported here resulted in luciferase expression 100 times higher than the levels obtained with previously described protocols. To verify whether sequences containing different trans-splicing signals influence reporter gene expression, we compared DNA fragments corresponding to 5′ untranslated plus intergenic (5′ UTR plus Ig) regions from GAPDH, TcP2β, α- and β-tubulin and amastin genes. Vectors containing sequences derived from the first four genes presented similar efficiencies and resulted in luciferase expression in transiently transfected epimastigotes that was up to 10 times higher than that for a control vector. In contrast, the amastin 5′ UTR plus Ig resulted in lower levels of reporter gene expression. We also constructed a vector containing an expression cassette designed to be targeted to the tubulin locus of the parasite.

Registro:

Documento: Artículo
Título:Expression of exogenous genes in Trypanosoma cruzi: Improving vectors and electroporation protocols
Autor:DaRocha, W.D.; Silva, R.A.; Bartholomeu, D.C.; Pires, S.F.; Freitas, J.M.; Macedo, A.M.; Vazquez, M.P.; Levin, M.J.; Teixeira, S.M.R.
Filiación:Depto. de Bioquim. e Imunologia, ICB, Universidade Federal de Minas Gerais, MG 31270-010 Belo Horizonte, Brazil
Lab. Biol. Molec. Enfermedad Chagas, School of Sciences, University of Buenos Aires, 1428 Buenos Aires, Argentina
Palabras clave:alpha tubulin; beta tubulin; DNA fragment; immunoglobulin; luciferase; red fluorescent protein; ribosome protein; 5' untranslated region; article; electroporation; epimastigote; expression vector; gene expression; microbial genetics; nonhuman; priority journal; reporter gene; Trypanosoma cruzi; 5' Untranslated Regions; Animals; Base Sequence; Electroporation; Genes, Reporter; Genetic Vectors; Green Fluorescent Proteins; Luciferases; Luminescent Proteins; Molecular Sequence Data; Protozoan Proteins; Transfection; Trypanosoma cruzi; Tubulin; Protozoa; Trypanosoma; Trypanosoma cruzi
Año:2004
Volumen:92
Número:2
Página de inicio:113
Página de fin:120
DOI: http://dx.doi.org/10.1007/s00436-003-1004-5
Título revista:Parasitology Research
Título revista abreviado:Parasitol. Res.
ISSN:09320113
CODEN:PARRE
CAS:alpha tubulin, 78769-62-7; beta tubulin, 87090-36-6; immunoglobulin, 9007-83-4; red fluorescent protein, 251925-26-5; 5' Untranslated Regions; Green Fluorescent Proteins, 147336-22-9; Luciferases, EC 1.13.12.-; Luminescent Proteins; Protozoan Proteins; red fluorescent protein; Tubulin
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_09320113_v92_n2_p113_DaRocha

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Citas:

---------- APA ----------
DaRocha, W.D., Silva, R.A., Bartholomeu, D.C., Pires, S.F., Freitas, J.M., Macedo, A.M., Vazquez, M.P.,..., Teixeira, S.M.R. (2004) . Expression of exogenous genes in Trypanosoma cruzi: Improving vectors and electroporation protocols. Parasitology Research, 92(2), 113-120.
http://dx.doi.org/10.1007/s00436-003-1004-5
---------- CHICAGO ----------
DaRocha, W.D., Silva, R.A., Bartholomeu, D.C., Pires, S.F., Freitas, J.M., Macedo, A.M., et al. "Expression of exogenous genes in Trypanosoma cruzi: Improving vectors and electroporation protocols" . Parasitology Research 92, no. 2 (2004) : 113-120.
http://dx.doi.org/10.1007/s00436-003-1004-5
---------- MLA ----------
DaRocha, W.D., Silva, R.A., Bartholomeu, D.C., Pires, S.F., Freitas, J.M., Macedo, A.M., et al. "Expression of exogenous genes in Trypanosoma cruzi: Improving vectors and electroporation protocols" . Parasitology Research, vol. 92, no. 2, 2004, pp. 113-120.
http://dx.doi.org/10.1007/s00436-003-1004-5
---------- VANCOUVER ----------
DaRocha, W.D., Silva, R.A., Bartholomeu, D.C., Pires, S.F., Freitas, J.M., Macedo, A.M., et al. Expression of exogenous genes in Trypanosoma cruzi: Improving vectors and electroporation protocols. Parasitol. Res. 2004;92(2):113-120.
http://dx.doi.org/10.1007/s00436-003-1004-5