Expression and maintenance of plasmid resistance in regenerating protoplasts of Bacillus subtilis

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Expression of plasmid-encoded resistances in regenerating protoplasts of Bacillus subtilis occurs only after wall synthesis has been resumed. This is observed for protoplasts obtained from cells already containing plasmids (pC194, pT127, pK545) or for plasmid-bearing cells coming from a PEG-mediated...

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Autor principal: Sanchez-Rivas, C.
Formato: JOUR
Materias:
Bacillus subtilis, Plasmid, Protoplast
Transformation, Gene expression, Cell wall regeneration
Acceso en línea:http://hdl.handle.net/20.500.12110/paper_09232508_v139_n4_p403_SanchezRivas
Aporte de:
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires
id todo:paper_09232508_v139_n4_p403_SanchezRivas
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spelling todo:paper_09232508_v139_n4_p403_SanchezRivas2023-10-03T15:45:51Z Expression and maintenance of plasmid resistance in regenerating protoplasts of Bacillus subtilis Sanchez-Rivas, C. Bacillus subtilis, Plasmid, Protoplast Transformation, Gene expression, Cell wall regeneration Expression of plasmid-encoded resistances in regenerating protoplasts of Bacillus subtilis occurs only after wall synthesis has been resumed. This is observed for protoplasts obtained from cells already containing plasmids (pC194, pT127, pK545) or for plasmid-bearing cells coming from a PEG-mediated transformation. Recovery of expression needs a 2-h incubation of protoplasts, previously washed to get rid of their lysozyme content, in rich hypertonic medium (SMMP). A longer incubation (24-h) results in the obtention of regenerants; however, most of them have lost their resistant phenotype in contrast to those obtained from the usual solid regeneration plates. This finding suggests either a high curing effect or some kind of gene inactivation phenomenon. Discussion is focused on the critical points that have to be considered when polyethylenglycol-mediated transformation of protoplasts is applied to recombinant DNA technology. © 1988. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_09232508_v139_n4_p403_SanchezRivas
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-134
collection Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
topic Bacillus subtilis, Plasmid, Protoplast
Transformation, Gene expression, Cell wall regeneration
spellingShingle Bacillus subtilis, Plasmid, Protoplast
Transformation, Gene expression, Cell wall regeneration
Sanchez-Rivas, C.
Expression and maintenance of plasmid resistance in regenerating protoplasts of Bacillus subtilis
topic_facet Bacillus subtilis, Plasmid, Protoplast
Transformation, Gene expression, Cell wall regeneration
description Expression of plasmid-encoded resistances in regenerating protoplasts of Bacillus subtilis occurs only after wall synthesis has been resumed. This is observed for protoplasts obtained from cells already containing plasmids (pC194, pT127, pK545) or for plasmid-bearing cells coming from a PEG-mediated transformation. Recovery of expression needs a 2-h incubation of protoplasts, previously washed to get rid of their lysozyme content, in rich hypertonic medium (SMMP). A longer incubation (24-h) results in the obtention of regenerants; however, most of them have lost their resistant phenotype in contrast to those obtained from the usual solid regeneration plates. This finding suggests either a high curing effect or some kind of gene inactivation phenomenon. Discussion is focused on the critical points that have to be considered when polyethylenglycol-mediated transformation of protoplasts is applied to recombinant DNA technology. © 1988.
format JOUR
author Sanchez-Rivas, C.
author_facet Sanchez-Rivas, C.
author_sort Sanchez-Rivas, C.
title Expression and maintenance of plasmid resistance in regenerating protoplasts of Bacillus subtilis
title_short Expression and maintenance of plasmid resistance in regenerating protoplasts of Bacillus subtilis
title_full Expression and maintenance of plasmid resistance in regenerating protoplasts of Bacillus subtilis
title_fullStr Expression and maintenance of plasmid resistance in regenerating protoplasts of Bacillus subtilis
title_full_unstemmed Expression and maintenance of plasmid resistance in regenerating protoplasts of Bacillus subtilis
title_sort expression and maintenance of plasmid resistance in regenerating protoplasts of bacillus subtilis
url http://hdl.handle.net/20.500.12110/paper_09232508_v139_n4_p403_SanchezRivas
work_keys_str_mv AT sanchezrivasc expressionandmaintenanceofplasmidresistanceinregeneratingprotoplastsofbacillussubtilis
_version_ 1807317281865203712

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