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Registro Completo |
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Biblioteca(s): |
Embrapa Arroz e Feijão. |
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Data corrente: |
30/09/2014 |
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Data da última atualização: |
26/02/2015 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
VENTURELLI, G. L.; BROD, F. C. A.; ROSSI, G. B.; ZIMMERMANN, N. F.; OLIVEIRA, J. P.; FARIA, J. C.; ARISI, A. C. M. |
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Afiliação: |
GUSTAVO L. VENTURELLI, UNIVERSIDADE FEDERAL DE SANTA CATARINA; FÁBIO C. A. BROD, UNIVERSIDADE FEDERAL DE SANTA CATARINA; GABRIELA B. ROSSI, UNIVERSIDADE FEDERAL DE SANTA CATARINA; NAIRA F. ZIMMERMANN, UNIVERSIDADE FEDERAL DE SANTA CATARINA; JAISON PEREIRA DE OLIVEIRA, CNPAF; JOSIAS CORREA DE FARIA, CNPAF; ANA C. M. ARISI, UNIVERSIDADE FEDERAL DE SANTA CATARINA. |
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Título: |
A specific endogenous reference for genetically modified common bean (Phaseolus vulgaris L.) DNA quantification by real-time PCR targeting lectin gene. |
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Ano de publicação: |
2014 |
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Fonte/Imprenta: |
Molecular Biotechnology, Totowa, v. 56, n. 11, p. 1060-1068, Nov. 2014. |
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DOI: |
10.1007/s12033-014-9786-5 |
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Idioma: |
Inglês |
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Conteúdo: |
The Embrapa 5.1 genetically modified (GM) common bean was approved for commercialization in Brazil. Methods for the quantification of this new genetically modified organism (GMO) are necessary. The development of a suitable endogenous reference is essential for GMO quantification by real-time PCR. Based on this, a new taxon-specific endogenous reference quantification assay was developed for Phaseolus vulgaris L. Three genes encoding common bean proteins (phaseolin, arcelin, and lectin) were selected as candidates for endogenous reference. Primers targeting these candidate genes were designed and the detection was evaluated using the SYBR Green chemistry. The assay targeting lectin gene showed higher specificity than the remaining assays, and a hydrolysis probe was then designed. This assay showed high specificity for 50 common bean samples from two gene pools, Andean and Mesoamerican. For GM common bean varieties, the results were similar to those obtained for non-GM isogenic varieties with PCR efficiency values ranging from 92 to 101 %. Moreover, this assay presented a limit of detection of ten haploid genome copies. The primers and probe developed in this work are suitable to detect and quantify either GM or non-GM common bean. |
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Thesagro: |
DNA; Engenharia genética; Feijão; Organismo transgênico; Phaseolus vulgaris; Planta transgênica. |
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Categoria do assunto: |
X Pesquisa, Tecnologia e Engenharia |
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Marc: |
LEADER 02148naa a2200277 a 4500
001 1996199
005 2015-02-26
008 2014 bl uuuu u00u1 u #d
024 7 $a10.1007/s12033-014-9786-5$2DOI
100 1 $aVENTURELLI, G. L.
245 $aA specific endogenous reference for genetically modified common bean (Phaseolus vulgaris L.) DNA quantification by real-time PCR targeting lectin gene.$h[electronic resource]
260 $c2014
520 $aThe Embrapa 5.1 genetically modified (GM) common bean was approved for commercialization in Brazil. Methods for the quantification of this new genetically modified organism (GMO) are necessary. The development of a suitable endogenous reference is essential for GMO quantification by real-time PCR. Based on this, a new taxon-specific endogenous reference quantification assay was developed for Phaseolus vulgaris L. Three genes encoding common bean proteins (phaseolin, arcelin, and lectin) were selected as candidates for endogenous reference. Primers targeting these candidate genes were designed and the detection was evaluated using the SYBR Green chemistry. The assay targeting lectin gene showed higher specificity than the remaining assays, and a hydrolysis probe was then designed. This assay showed high specificity for 50 common bean samples from two gene pools, Andean and Mesoamerican. For GM common bean varieties, the results were similar to those obtained for non-GM isogenic varieties with PCR efficiency values ranging from 92 to 101 %. Moreover, this assay presented a limit of detection of ten haploid genome copies. The primers and probe developed in this work are suitable to detect and quantify either GM or non-GM common bean.
650 $aDNA
650 $aEngenharia genética
650 $aFeijão
650 $aOrganismo transgênico
650 $aPhaseolus vulgaris
650 $aPlanta transgênica
700 1 $aBROD, F. C. A.
700 1 $aROSSI, G. B.
700 1 $aZIMMERMANN, N. F.
700 1 $aOLIVEIRA, J. P.
700 1 $aFARIA, J. C.
700 1 $aARISI, A. C. M.
773 $tMolecular Biotechnology, Totowa$gv. 56, n. 11, p. 1060-1068, Nov. 2014.
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Registro original: |
Embrapa Arroz e Feijão (CNPAF) |
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