Protoplast Transformation of Recalcitrant Alkaliphilic Bacillus sp. with Methylated Plasmid DNA and a Developed Hard Agar Regeneration Medium

Among the diverse alkaliphilic Bacillus strains, only a little have been reported to be genetically transformed. In this study, an efficient protoplast transformation procedure was developed for recalcitrant alkaliphilic Bacillus sp. N16-5. The procedure involved polyethylene glycol-induced DNA upta...

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Detalles Bibliográficos
Autores principales: Gao, Chenghua, Xue, Yanfen, Ma, Yanhe
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2011
Materias:
Research Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3223237/
https://www.ncbi.nlm.nih.gov/pubmed/22132233
http://dx.doi.org/10.1371/journal.pone.0028148
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author Gao, Chenghua
Xue, Yanfen
Ma, Yanhe
author_facet Gao, Chenghua
Xue, Yanfen
Ma, Yanhe
author_sort Gao, Chenghua
collection PubMed
description Among the diverse alkaliphilic Bacillus strains, only a little have been reported to be genetically transformed. In this study, an efficient protoplast transformation procedure was developed for recalcitrant alkaliphilic Bacillus sp. N16-5. The procedure involved polyethylene glycol-induced DNA uptake by the protoplasts and subsequent protoplast regeneration with a developed hard agar regeneration medium. An in vivo methylation strategy was introduced to methylate the exogenous plasmid DNA for improving the transformation efficiency. The transformation efficiency reached to 1.1×10(5) transformants per µg plasmid DNA with methylated plasmid pHCMC04 and the developed hard agar regeneration medium. This procedure might also be applicable to the genetic transformation of other Bacillus strains.
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spelling pubmed-32232372011-11-30 Protoplast Transformation of Recalcitrant Alkaliphilic Bacillus sp. with Methylated Plasmid DNA and a Developed Hard Agar Regeneration Medium Gao, Chenghua Xue, Yanfen Ma, Yanhe PLoS One Research Article Among the diverse alkaliphilic Bacillus strains, only a little have been reported to be genetically transformed. In this study, an efficient protoplast transformation procedure was developed for recalcitrant alkaliphilic Bacillus sp. N16-5. The procedure involved polyethylene glycol-induced DNA uptake by the protoplasts and subsequent protoplast regeneration with a developed hard agar regeneration medium. An in vivo methylation strategy was introduced to methylate the exogenous plasmid DNA for improving the transformation efficiency. The transformation efficiency reached to 1.1×10(5) transformants per µg plasmid DNA with methylated plasmid pHCMC04 and the developed hard agar regeneration medium. This procedure might also be applicable to the genetic transformation of other Bacillus strains. Public Library of Science 2011-11-23 /pmc/articles/PMC3223237/ /pubmed/22132233 http://dx.doi.org/10.1371/journal.pone.0028148 Text en Gao et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Gao, Chenghua
Xue, Yanfen
Ma, Yanhe
Protoplast Transformation of Recalcitrant Alkaliphilic Bacillus sp. with Methylated Plasmid DNA and a Developed Hard Agar Regeneration Medium
title Protoplast Transformation of Recalcitrant Alkaliphilic Bacillus sp. with Methylated Plasmid DNA and a Developed Hard Agar Regeneration Medium
title_full Protoplast Transformation of Recalcitrant Alkaliphilic Bacillus sp. with Methylated Plasmid DNA and a Developed Hard Agar Regeneration Medium
title_fullStr Protoplast Transformation of Recalcitrant Alkaliphilic Bacillus sp. with Methylated Plasmid DNA and a Developed Hard Agar Regeneration Medium
title_full_unstemmed Protoplast Transformation of Recalcitrant Alkaliphilic Bacillus sp. with Methylated Plasmid DNA and a Developed Hard Agar Regeneration Medium
title_short Protoplast Transformation of Recalcitrant Alkaliphilic Bacillus sp. with Methylated Plasmid DNA and a Developed Hard Agar Regeneration Medium
title_sort protoplast transformation of recalcitrant alkaliphilic bacillus sp. with methylated plasmid dna and a developed hard agar regeneration medium
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3223237/
https://www.ncbi.nlm.nih.gov/pubmed/22132233
http://dx.doi.org/10.1371/journal.pone.0028148
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AT mayanhe protoplasttransformationofrecalcitrantalkaliphilicbacillusspwithmethylatedplasmiddnaandadevelopedhardagarregenerationmedium

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