A simple, economical and reproducible protein extraction protocol for proteomics studies of soybean roots

Detalhes bibliográficos
Autor(a) principal: Rodrigues,Elisete Pains
Data de Publicação: 2012
Outros Autores: Torres,Adalgisa Ribeiro, Batista,Jesiane Stefânia da Silva, Huergo,Luciano, Hungria,Mariangela
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Genetics and Molecular Biology
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572012000200016
Resumo: Sample preparation is a critical step in two-dimensional gel electrophoresis (2-DE) of plant tissues. Here we describe a phenol/SDS procedure that, although greatly simplified, produced well-resolved and reproducible 2-DE profiles of protein extracts from soybean [Glycine max (L.) Merril] roots. Extractions were made in three replicates using both the original and simplified procedure. To evaluate the quality of the extracted proteins, ten spots were randomly selected and identified by mass spectrometry (MS). The 2-DE gels were equally well resolved, with no streaks or smears, and no significant differences were observed in protein yield, reproducibility, resolution or number of spots. Mass spectra of the ten selected spots were compared with database entries and allowed high-quality identification of proteins. The simplified protocol described here presents considerable savings of time and reagents without compromising the quality of 2-DE protein profiles and compatibility with MS analysis, and may facilitate the progress of proteomics studies of legume-rhizobia interactions.
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spelling A simple, economical and reproducible protein extraction protocol for proteomics studies of soybean roots2-DEprotein extractionproteomicssoybean rootssymbiosisSample preparation is a critical step in two-dimensional gel electrophoresis (2-DE) of plant tissues. Here we describe a phenol/SDS procedure that, although greatly simplified, produced well-resolved and reproducible 2-DE profiles of protein extracts from soybean [Glycine max (L.) Merril] roots. Extractions were made in three replicates using both the original and simplified procedure. To evaluate the quality of the extracted proteins, ten spots were randomly selected and identified by mass spectrometry (MS). The 2-DE gels were equally well resolved, with no streaks or smears, and no significant differences were observed in protein yield, reproducibility, resolution or number of spots. Mass spectra of the ten selected spots were compared with database entries and allowed high-quality identification of proteins. The simplified protocol described here presents considerable savings of time and reagents without compromising the quality of 2-DE protein profiles and compatibility with MS analysis, and may facilitate the progress of proteomics studies of legume-rhizobia interactions.Sociedade Brasileira de Genética2012-01-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572012000200016Genetics and Molecular Biology v.35 n.1 suppl.1 2012reponame:Genetics and Molecular Biologyinstname:Sociedade Brasileira de Genética (SBG)instacron:SBG10.1590/S1415-47572012000200016info:eu-repo/semantics/openAccessRodrigues,Elisete PainsTorres,Adalgisa RibeiroBatista,Jesiane Stefânia da SilvaHuergo,LucianoHungria,Mariangelaeng2012-06-01T00:00:00Zoai:scielo:S1415-47572012000200016Revistahttp://www.gmb.org.br/ONGhttps://old.scielo.br/oai/scielo-oai.php||editor@gmb.org.br1678-46851415-4757opendoar:2012-06-01T00:00Genetics and Molecular Biology - Sociedade Brasileira de Genética (SBG)false
dc.title.none.fl_str_mv A simple, economical and reproducible protein extraction protocol for proteomics studies of soybean roots
title A simple, economical and reproducible protein extraction protocol for proteomics studies of soybean roots
spellingShingle A simple, economical and reproducible protein extraction protocol for proteomics studies of soybean roots
Rodrigues,Elisete Pains
2-DE
protein extraction
proteomics
soybean roots
symbiosis
title_short A simple, economical and reproducible protein extraction protocol for proteomics studies of soybean roots
title_full A simple, economical and reproducible protein extraction protocol for proteomics studies of soybean roots
title_fullStr A simple, economical and reproducible protein extraction protocol for proteomics studies of soybean roots
title_full_unstemmed A simple, economical and reproducible protein extraction protocol for proteomics studies of soybean roots
title_sort A simple, economical and reproducible protein extraction protocol for proteomics studies of soybean roots
author Rodrigues,Elisete Pains
author_facet Rodrigues,Elisete Pains
Torres,Adalgisa Ribeiro
Batista,Jesiane Stefânia da Silva
Huergo,Luciano
Hungria,Mariangela
author_role author
author2 Torres,Adalgisa Ribeiro
Batista,Jesiane Stefânia da Silva
Huergo,Luciano
Hungria,Mariangela
author2_role author
author
author
author
dc.contributor.author.fl_str_mv Rodrigues,Elisete Pains
Torres,Adalgisa Ribeiro
Batista,Jesiane Stefânia da Silva
Huergo,Luciano
Hungria,Mariangela
dc.subject.por.fl_str_mv 2-DE
protein extraction
proteomics
soybean roots
symbiosis
topic 2-DE
protein extraction
proteomics
soybean roots
symbiosis
description Sample preparation is a critical step in two-dimensional gel electrophoresis (2-DE) of plant tissues. Here we describe a phenol/SDS procedure that, although greatly simplified, produced well-resolved and reproducible 2-DE profiles of protein extracts from soybean [Glycine max (L.) Merril] roots. Extractions were made in three replicates using both the original and simplified procedure. To evaluate the quality of the extracted proteins, ten spots were randomly selected and identified by mass spectrometry (MS). The 2-DE gels were equally well resolved, with no streaks or smears, and no significant differences were observed in protein yield, reproducibility, resolution or number of spots. Mass spectra of the ten selected spots were compared with database entries and allowed high-quality identification of proteins. The simplified protocol described here presents considerable savings of time and reagents without compromising the quality of 2-DE protein profiles and compatibility with MS analysis, and may facilitate the progress of proteomics studies of legume-rhizobia interactions.
publishDate 2012
dc.date.none.fl_str_mv 2012-01-01
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572012000200016
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572012000200016
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1590/S1415-47572012000200016
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv text/html
dc.publisher.none.fl_str_mv Sociedade Brasileira de Genética
publisher.none.fl_str_mv Sociedade Brasileira de Genética
dc.source.none.fl_str_mv Genetics and Molecular Biology v.35 n.1 suppl.1 2012
reponame:Genetics and Molecular Biology
instname:Sociedade Brasileira de Genética (SBG)
instacron:SBG
instname_str Sociedade Brasileira de Genética (SBG)
instacron_str SBG
institution SBG
reponame_str Genetics and Molecular Biology
collection Genetics and Molecular Biology
repository.name.fl_str_mv Genetics and Molecular Biology - Sociedade Brasileira de Genética (SBG)
repository.mail.fl_str_mv ||editor@gmb.org.br
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