Artículo
Abstract:
Single strand conformation analysis (SSCA) is a technique that has been used to detect point mutations. We explored its usefulness in the analysis of four different members of the Trypanosoma cruzi TcP2β gene family and its suitability for detection of polymorphism in different parasite strains. The availability of primers covering a 97-bp sequence at the 5' end of the genes allowed assessment of the effect of a single base substitution, while the analysis of a 321 bp long sequence permitted the evaluation of sequences differing in several bases. PCR products were analysed under four different electrophoretic conditions: with or without the addition of 10% glycerol in a 6% polyacrylamide gel run at room temperature or at 4°C. Shifts in mobility were radically dependent on the migration condition. Both 97-bp and 321-bp amplicons were best resolved at 4°C, without glycerol. Amplification products derived from total genomic DNA showed a pattern that resembled closely a combination of the products derived from the cloned genes. The results herein demonstrate the usefulness of SSCA to differentiate forms of a complex protozoan gene family, and to scan its polymorphic nature. Furthermore, due to the remarkable sensitivity of the technique it can generate genomic markers, such as Sequence Tagged Sites (STS), of great need in the T. cruzi genome project.
Registro:
| Documento: | Artículo |
| Título: | Detection of polymorphism in the Trypanosoma cruzi TcP2β gene family by single strand conformational analysis (SSCA) |
| Autor: | Vázquez, M.P.; Beldjord, C.; Lorenzi, H.; Bienvenu, T.; Levín, M.J. |
| Filiación: | Inst. de Invest. en Ingenieria G., Vuelta de Obligado 2490 2P, 1428, Buenos Aires, Argentina Service de Biochimie Genetique, Hôpital Cochin, Paris, France |
| Palabras clave: | Protozoan parasites genome projects; Ribosomal P protein genes; Sequence Tagged Sites; Trypanosoma cruzi; glycerol; glycine cleavage system; polyacrylamide gel; ribosome protein; article; controlled study; dna polymorphism; electrophoretic mobility; gene amplification; genome; marker gene; multigene family; nonhuman; nucleic acid base substitution; point mutation; polymerase chain reaction; priority journal; single strand conformation polymorphism; trypanosoma cruzi; Animals; Base Sequence; Genes, Protozoan; Molecular Sequence Data; Multigene Family; Phosphoproteins; Polymerase Chain Reaction; Polymorphism, Single-Stranded Conformational; Protozoan Proteins; Ribosomal Proteins; Sequence Tagged Sites; Trypanosoma cruzi; Protozoa; Trypanosoma; Trypanosoma cruzi |
| Año: | 1996 |
| Volumen: | 180 |
| Número: | 1-2 |
| Página de inicio: | 43 |
| Página de fin: | 48 |
| DOI: | http://dx.doi.org/10.1016/S0378-1119(96)00401-5 |
| Título revista: | Gene |
| Título revista abreviado: | GENE |
| ISSN: | 03781119 |
| CODEN: | GENED |
| CAS: | phosphoprotein P2, ribosomal; Phosphoproteins; Protozoan Proteins; Ribosomal Proteins |
| Registro: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_03781119_v180_n1-2_p43_Vazquez |
Referencias:
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Citas:
---------- APA ----------
Vázquez, M.P., Beldjord, C., Lorenzi, H., Bienvenu, T. & Levín, M.J. (1996) . Detection of polymorphism in the Trypanosoma cruzi TcP2β gene family by single strand conformational analysis (SSCA). Gene, 180(1-2), 43-48.http://dx.doi.org/10.1016/S0378-1119(96)00401-5
---------- CHICAGO ----------
Vázquez, M.P., Beldjord, C., Lorenzi, H., Bienvenu, T., Levín, M.J. "Detection of polymorphism in the Trypanosoma cruzi TcP2β gene family by single strand conformational analysis (SSCA)" . Gene 180, no. 1-2 (1996) : 43-48.http://dx.doi.org/10.1016/S0378-1119(96)00401-5
---------- MLA ----------
Vázquez, M.P., Beldjord, C., Lorenzi, H., Bienvenu, T., Levín, M.J. "Detection of polymorphism in the Trypanosoma cruzi TcP2β gene family by single strand conformational analysis (SSCA)" . Gene, vol. 180, no. 1-2, 1996, pp. 43-48.http://dx.doi.org/10.1016/S0378-1119(96)00401-5
---------- VANCOUVER ----------
Vázquez, M.P., Beldjord, C., Lorenzi, H., Bienvenu, T., Levín, M.J. Detection of polymorphism in the Trypanosoma cruzi TcP2β gene family by single strand conformational analysis (SSCA). GENE. 1996;180(1-2):43-48.http://dx.doi.org/10.1016/S0378-1119(96)00401-5
