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 | Acceso al texto completo restringido a Biblioteca INIA Las Brujas. Por información adicional contacte bibliolb@inia.org.uy. |
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Registro completo
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Biblioteca (s) : |
INIA Las Brujas. |
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Fecha : |
06/05/2021 |
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Actualizado : |
10/08/2021 |
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Tipo de producción científica : |
Artículos en Revistas Indexadas Internacionales |
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Autor : |
NARANCIO, R.; JOHN, U.; MASON, J.; GIRALDO, P.; SPANGENBERG, G. |
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Afiliación : |
RAFAEL NARANCIO FERES, INIA (Instituto Nacional de Investigación Agropecuaria), Uruguay; School of Applied Systems Biology, La Trobe University, Melbourne, VIC, Australia; Agriculture Victoria Research, AgriBio, Centre for AgriBioscience, Melbourne, VIC, Australia; ULRIK JOHN, Agriculture Victoria Research, AgriBio, Centre for AgriBioscience, Melbourne, VIC, Australia; JOHN MASON, School of Applied Systems Biology, La Trobe University, Melbourne, VIC, Australia; Agriculture Victoria Research, AgriBio, Centre for AgriBioscience, Melbourne, VIC, Australia; PAULA GIRALDO, Agriculture Victoria Research, AgriBio, Centre for AgriBioscience, Melbourne, VIC, Australia; Faculty of Veterinary and Agricultural Sciences, The University of Melbourne, Melbourne, VIC, Australia; GERMAN SPANGENBERG, School of Applied Systems Biology, La Trobe University, Melbourne, VIC, Australia; Agriculture Victoria Research, AgriBio, Centre for AgriBioscience, Melbourne, VIC, Australia. |
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Título : |
Digital PCR (dPCR) and qPCR mediated determination of transgene copy number in the forage legume white clover (Trifolium repens). |
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Fecha de publicación : |
2021 |
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Fuente / Imprenta : |
Molecular Biology Reports, 2021, volume 48, Issue 4, Pages 3069 - 3077. Doi: https://doi.org/10.1007/s11033-021-06354-5 |
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ISSN : |
0301-4851 |
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DOI : |
10.1007/s11033-021-06354-5 |
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Idioma : |
Inglés |
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Notas : |
Article history: Received, 03 February 2021; Accepted, 12 April 2021; Published, 16 April 2021.
This project was funded by Agriculture Victoria, and Dairy Australia. |
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Contenido : |
ABSTRACT.
Obtaining data on transgene copy number is an integral step in the generation of transgenic plants. Techniques such as Southern blot, segregation analysis, and quantitative PCR (qPCR) have routinely been used for this task, in a range of species. More recently, use of Digital PCR (dPCR) has become prevalent, with a measurement accuracy higher than qPCR reported. Here, the relative merits of qPCR and dPCR for transgene copy number estimation in white clover were investigated. Furthermore, given that single copy reference genes are desirable for estimating gene copy number by relative quantification, and that no single-copy genes have been reported in this species, a search and evaluation of suitable reference genes in white clover was undertaken. Results demonstrated a higher accuracy of dPCR relative to qPCR for copy number estimation in white clover. Two genes, Pyruvate dehydrogenase (PDH), and an ATP-dependent protease, identified as single-copy genes, were used as references for copy number estimation by relative quantification. Identification of single-copy genes in white clover will enable the application of relative quantification for copy number estimation of other genes or transgenes in the species. The results generated here validate the use of dPCR as a reliable strategy for transgene copy number estimation in white clover, and provide resources for future copy number studies in this species.
© 2021, The Author(s), under exclusive licence to Springer Nature B.V. MenosABSTRACT.
Obtaining data on transgene copy number is an integral step in the generation of transgenic plants. Techniques such as Southern blot, segregation analysis, and quantitative PCR (qPCR) have routinely been used for this task, in a range of species. More recently, use of Digital PCR (dPCR) has become prevalent, with a measurement accuracy higher than qPCR reported. Here, the relative merits of qPCR and dPCR for transgene copy number estimation in white clover were investigated. Furthermore, given that single copy reference genes are desirable for estimating gene copy number by relative quantification, and that no single-copy genes have been reported in this species, a search and evaluation of suitable reference genes in white clover was undertaken. Results demonstrated a higher accuracy of dPCR relative to qPCR for copy number estimation in white clover. Two genes, Pyruvate dehydrogenase (PDH), and an ATP-dependent protease, identified as single-copy genes, were used as references for copy number estimation by relative quantification. Identification of single-copy genes in white clover will enable the application of relative quantification for copy number estimation of other genes or transgenes in the species. The results generated here validate the use of dPCR as a reliable strategy for transgene copy number estimation in white clover, and provide resources for future copy number studies in this species.
© 2021, The Author(s), under exclusive licence to Springer Nat... Presentar Todo |
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Palabras claves : |
DdPCR; Reference genes; Relative quantification; Single copy genes; Trifolium repens; Zygosity determination. |
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Asunto categoría : |
F30 Genética vegetal y fitomejoramiento |
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Marc : |
LEADER 02569naa a2200277 a 4500
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024 7 $a10.1007/s11033-021-06354-5$2DOI
100 1 $aNARANCIO, R.
245 $aDigital PCR (dPCR) and qPCR mediated determination of transgene copy number in the forage legume white clover (Trifolium repens).$h[electronic resource]
260 $c2021
500 $aArticle history: Received, 03 February 2021; Accepted, 12 April 2021; Published, 16 April 2021. This project was funded by Agriculture Victoria, and Dairy Australia.
520 $aABSTRACT. Obtaining data on transgene copy number is an integral step in the generation of transgenic plants. Techniques such as Southern blot, segregation analysis, and quantitative PCR (qPCR) have routinely been used for this task, in a range of species. More recently, use of Digital PCR (dPCR) has become prevalent, with a measurement accuracy higher than qPCR reported. Here, the relative merits of qPCR and dPCR for transgene copy number estimation in white clover were investigated. Furthermore, given that single copy reference genes are desirable for estimating gene copy number by relative quantification, and that no single-copy genes have been reported in this species, a search and evaluation of suitable reference genes in white clover was undertaken. Results demonstrated a higher accuracy of dPCR relative to qPCR for copy number estimation in white clover. Two genes, Pyruvate dehydrogenase (PDH), and an ATP-dependent protease, identified as single-copy genes, were used as references for copy number estimation by relative quantification. Identification of single-copy genes in white clover will enable the application of relative quantification for copy number estimation of other genes or transgenes in the species. The results generated here validate the use of dPCR as a reliable strategy for transgene copy number estimation in white clover, and provide resources for future copy number studies in this species. © 2021, The Author(s), under exclusive licence to Springer Nature B.V.
653 $aDdPCR
653 $aReference genes
653 $aRelative quantification
653 $aSingle copy genes
653 $aTrifolium repens
653 $aZygosity determination
700 1 $aJOHN, U.
700 1 $aMASON, J.
700 1 $aGIRALDO, P.
700 1 $aSPANGENBERG, G.
773 $tMolecular Biology Reports, 2021, volume 48, Issue 4, Pages 3069 - 3077. Doi: https://doi.org/10.1007/s11033-021-06354-5
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Registro original : |
INIA Las Brujas (LB) |
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| Registros recuperados : 13 | |
| 3. |  | NARANCIO, R.; JOHN, U.; MASON, J.; GIRALDO, P.; SPANGENBERG, G. Digital PCR (dPCR) and qPCR mediated determination of transgene copy number in the forage legume white clover (Trifolium repens). Molecular Biology Reports, 2021, volume 48, Issue 4, Pages 3069 - 3077. Doi: https://doi.org/10.1007/s11033-021-06354-5 Article history: Received, 03 February 2021; Accepted, 12 April 2021; Published, 16 April 2021.
This project was funded by Agriculture Victoria, and Dairy Australia.| Tipo: Artículos en Revistas Indexadas Internacionales | Circulación / Nivel : Internacional - -- |
| Biblioteca(s): INIA Las Brujas. |
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| 4. | | NARANCIO, R.; JOHN, U.; MASON, J.; SPANGENBERG, G. Selection of optimal reference genes for quantitative RT-PCR transcript abundance analysis in white clover (Trifolium repens L.). Functional Plant Biology, 2018, Volume 45, Issue 7, Pages 737-744. Doi: https://doi.org/10.1071/FP17304 Article history: Received 1 November 2017; Accepted 21 January 2018; Published online 16 February 2018.
Corresponding author: John, U.; Agriculture Victoria Research, AgriBio Centre for AgriBioscience, La Trobe University, Melbourne,...| Tipo: Artículos en Revistas Indexadas Internacionales | Circulación / Nivel : Internacional - -- |
| Biblioteca(s): INIA Las Brujas. |
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| 5. | | DALLA RIZZA, M.; DIAZ, P.; NARANCIO, R.; CABRERA, A.; FERREIRA, F. Biomolecules as host defense weapons against microbial pathogens. (Review) Recent Patents on DNA and Gene Sequences, 2008, v.2, no.2, p.82-96. Article history: Received: April 21, 2008 / Accepted: April 29, 2008 / Revised: April 29, 2008.| Tipo: Artículos en Revistas Indexadas Internacionales | Circulación / Nivel : Internacional - - |
| Biblioteca(s): INIA Las Brujas. |
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| 7. | | NARANCIO, R.; ZORRILLA, P.; ROBELLO, C.; GONZÁLEZ-ARCOS, M.; VILARÓ, F.; PRITSCH, C.; DALLA RIZZA, M. Insights on gene expression response of a characterized resistant genotype of Solanum commersonii Dun. against Ralstonia solanacearum. European Journal of Plant Pathology, 2013, v.136, no.4, p.823-835. Article history: Accepted: 27 March 2013 / Published online: 28 April 2013.| Tipo: Artículos en Revistas Indexadas Internacionales | Circulación / Nivel : B - 1 |
| Biblioteca(s): INIA Las Brujas. |
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| 8. | | ALEM, D.; NARANCIO, R.; DIAZ, P.; REBUFFO, M.; ZARZA, R.; DALLA RIZZA, M. Molecular characterization of Lotus corniculatus cultivars using transferable microsatellite markers. Ciencia e Investigación Agraria, Santiago de Chile, v. 38, n. 3, p. 463-471, 2011. Article history: Received September 7 2010. /Accepted June 21, 2011.| Tipo: Artículos en Revistas Indexadas Internacionales | Circulación / Nivel : B - 3 |
| Biblioteca(s): INIA La Estanzuela. |
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| 9. | | REAL, D.; REYNO, R.; DALLA RIZZA, M.; ALTIER, N.; DO CANTO, J.; NARANCIO, R.; FOLLE, G.; BURGUEÑO, J. Caracterización de genotipos de Paspalum notatum por tolerancia a bajas temperaturas y reacción frente a Claviceps paspali ln: Reunión del grupo técnico en forrajeras del Cono Sur, 22., 2008, Minas, Uruguay Bioma campos: innovando para mantener su sustentabilidad y competitividad. Memorias. Minas (Uruguay): INIA; FAO; PROCISUR, 2008. p. 156 Versión impresa y en CD ROM Instituto Nacional de Investigación Agropecuaria, Uruguay; FAO; PROCISUR| Tipo: Trabajos en Congresos/Conferencias |
| Biblioteca(s): INIA Las Brujas; INIA Tacuarembó. |
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| 10. | | NARANCIO, R.; DING, Y.L.; LIN, Y.H.; SAHAB, S.; PANTER, S.; HAYES, M.; JOHN, U.; MASON, J.; SPANGENBERG, G. Generación y caracterización molecular y funcional de plantas de trébol blanco transgénicas con las características de senescencia retardada, tolerancia a aluminio y resistencia a Alfalfa Mosaic Virus (AMV). [resumen] In: INIA (Instituto Nacional de Investigación Agropecuaria); INIA Las Brujas; Biotecnología. Jornada de Agrobiotecnología, XI. Encuentro Nacional de REDBIO, III. Jornada técnica. Las Brujas, Canelones (UY): INIA, 2018. p.10-11. (Serie Actividades de Difusión; 786)| Biblioteca(s): INIA Las Brujas. |
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| 11. | | NARANCIO, R.; DING, Y.-L.; LIN, Y.-H.; SAHAB, S.; PANTER, S.; HAYES, M.; JOHN, U.; ANDERSON, H.; MASON, J.; SPANGENBERG, G. Application of linked and unlinked co transformation to generate triple stack, marker free, transgenic white clover (Trifolium repens L.). Plant Cell, Tissue and Organ Culture, 2020. DOI: https://doi.org/10.1007/s11240-020-01891-6 Article history: Received: 19 May 2020 / Accepted: 15 July 2020.| Tipo: Artículos en Revistas Indexadas Internacionales | Circulación / Nivel : -- - -- |
| Biblioteca(s): INIA Las Brujas. |
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| 12. | | NARANCIO, R.; DING, Y.-L.; LIN, Y.-H.; SAHAB, S.; PANTER, S.; HAYES, M.; JOHN, U.; ANDERSON, H.; MASON, J.; SPANGENBERG, G. Correction to: Application of linked and unlinked co‑transformation to generate triple stack, marker‑free, transgenic white clover (Trifolium repens L.) (Plant Cell, Tissue and Organ Culture (PCTOC), (2020), 142, 3, (635-646), 10.1007/s11240-020-01891-6). Plant Cell, Tissue and Organ Culture, 2020. DOI: https://doi.org/10.1007/s11240-020-01891-6 Article history: Received: 19 May 2020 / Accepted: 15 July 2020.| Biblioteca(s): INIA Treinta y Tres. |
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| 13. | | REYNO, R.; NARANCIO, R.; SPERANZA, P.; DO CANTO, J.; LOPEZ-CARRO, B.; HERNANDEZ, P.; BURGUEÑO, J.; REAL, D.; DALLA RIZZA, M. Molecular and cytogenetic characterization of a collection of bahiagrass (Paspalum notatum Flügge) native to Uruguay. Genetic Resources and Crop Evolution, 2012, v. 59, no.8, p. 1823-1832. Article history: Received: 9 August 2011 / Accepted: 30 January 2012 / Published online: 22 February 2012.| Tipo: Artículos en Revistas Indexadas Internacionales | Circulación / Nivel : A - 2 |
| Biblioteca(s): INIA Las Brujas; INIA Tacuarembó. |
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| Registros recuperados : 13 | |
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