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 | Acesso ao texto completo restrito à biblioteca da Embrapa Recursos Genéticos e Biotecnologia. Para informações adicionais entre em contato com cenargen.biblioteca@embrapa.br. |
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Registro Completo |
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Biblioteca(s): |
Embrapa Recursos Genéticos e Biotecnologia. |
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Data corrente: |
26/08/2024 |
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Data da última atualização: |
26/08/2024 |
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Tipo da produção científica: |
Capítulo em Livro Técnico-Científico |
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Autoria: |
SILVA, M. S.; BERND, R. B.; SOUZA, A. L. X. de; MELO, L. A. M. P. de; PEREIRA, J. E. S. |
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Afiliação: |
MARILIA SANTOS SILVA PATRIOTA, CENARGEN; REGINA BEATRIZ BERND; ANDRE LUIS XAVIER DE SOUZA, CENARGEN; LUIS ALBERTO MARTINS P DE MELO, CENARGEN; JONNY EVERSON SCHERWINSKI PEREIRA, CENARGEN. |
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Título: |
Quantification of DNA methylation by ELISA in epigenetic studies in plant tissue culture: a theoretical-practical guide. |
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Ano de publicação: |
2024 |
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Fonte/Imprenta: |
In: Loyola-Vargas, V.; Ochoa-Alejo, N. (Ed.). Plant cell culture protocols. New York, NY: Humana, 2024. cap. 22, p. 323-350. (Methods in Molecular Biology), 2827). |
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DOI: |
10.1007/978-1-0716-3954-2_22 |
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Idioma: |
Inglês |
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Notas: |
Na publicação: Jonny Everson Scherwinski-Pereira. |
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Conteúdo: |
This chapter describes a step-by-step protocol for rapid serological quantification of global DNA methylation by enzyme-linked immunosorbent assay (ELISA) in plant tissue culture specimens. As a case study model, we used the coconut palm (Cocos nucifera), from which plumules were subjected to somatic embryogenesis followed by embryogenic calli multiplication. DNA methylation is one of the most common epigenetic markers in the regulation of gene expression. DNA methylation is generally associated with non-expressed genes, that is, gene silencing under certain conditions, and the degree of DNA methylation can be used as a marker of various physiological processes, both in plants and in animal cells. Methylation consists of adding a methyl radical to carbon 5 of the DNA cytosine base. Herein, the global DNA methylation was quantified by ELISA with antibodies against methylated cytosines using a commercial kit (Zymo-Research™). The method allowed the detection of methylation in total DNA extracts from coconut palm embryogenic calli (arising from somatic embryogenesis) cultivated in liquid or solid media by using antibodies against methylated cytosines and enzymatic development with a colorimetric substrate. Control samples of commercially provided Escherichia coli bacterial DNA with previously known methylation percentages were included in the ELISA test to construct an experimental methylation standard curve. The logarithmic regression of this E. coli standard curve allowed methylation quantification in coconut palm samples. The present ELISA methodology, applied to coconut palm tissue culture specimens, is promising for use in other plant species and botanical families. This chapter is presented in a suitable format for use as a step-by-step laboratory procedure manual, with theoretical introduction information, which makes it easy to apply the protocol in samples of any biological nature to evaluate DNA global methylation associated with any physiological process. MenosThis chapter describes a step-by-step protocol for rapid serological quantification of global DNA methylation by enzyme-linked immunosorbent assay (ELISA) in plant tissue culture specimens. As a case study model, we used the coconut palm (Cocos nucifera), from which plumules were subjected to somatic embryogenesis followed by embryogenic calli multiplication. DNA methylation is one of the most common epigenetic markers in the regulation of gene expression. DNA methylation is generally associated with non-expressed genes, that is, gene silencing under certain conditions, and the degree of DNA methylation can be used as a marker of various physiological processes, both in plants and in animal cells. Methylation consists of adding a methyl radical to carbon 5 of the DNA cytosine base. Herein, the global DNA methylation was quantified by ELISA with antibodies against methylated cytosines using a commercial kit (Zymo-Research™). The method allowed the detection of methylation in total DNA extracts from coconut palm embryogenic calli (arising from somatic embryogenesis) cultivated in liquid or solid media by using antibodies against methylated cytosines and enzymatic development with a colorimetric substrate. Control samples of commercially provided Escherichia coli bacterial DNA with previously known methylation percentages were included in the ELISA test to construct an experimental methylation standard curve. The logarithmic regression of this E. coli standard curve allowed met... Mostrar Tudo |
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Palavras-Chave: |
Plant tissue culture. |
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Thesagro: |
Elisa. |
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Thesaurus Nal: |
DNA methylation; Epigenetics. |
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Categoria do assunto: |
-- |
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Marc: |
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Registro original: |
Embrapa Recursos Genéticos e Biotecnologia (CENARGEN) |
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| 1. |  | RODRIGUES, T. M.; SILVA, D. F. da; NEVES, L.; GANOSA, G. A.; MORAES, L. F. D. de; LIMA, G. C. de; CHRISTO, A. G.; RIBEIRO, R. D.; FARIA, S. M. de. Legumiosae na Floresta Nacional de Carajás, Pará, Brasil. In: SIMPÓSIO NACIONAL SOBRE RECUPERAÇÃO DE ÁREAS DEGRADADAS, 9., 2012, Rio de Janeiro. Recriando ambientes sustentáveis: trabalhos voluntários. Rio de Janeiro: Sobrade, 2012.| Tipo: Artigo em Anais de Congresso |
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