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Abstract:

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.

Registro:

Documento: Artículo
Título:Expression and maintenance of plasmid resistance in regenerating protoplasts of Bacillus subtilis
Autor:Sanchez-Rivas, C.
Filiación:Instituto de Investigaciones en Ingenieia Genetica y Biologia Molecular (INGEBI-CONICET) Obligado 24, 1428 Buenos Aires, Argentina
Palabras clave:Bacillus subtilis, Plasmid, Protoplast; Transformation, Gene expression, Cell wall regeneration; bacillus subtilis; bacterium transformation; cell wall; gene expression; genetic engineering; nonhuman; plasmid; priority journal; protoplast; Bacillus subtilis; Cell Wall; Culture Media; Drug Resistance, Microbial; Gene Expression Regulation; Osmotic Pressure; Protoplasts; R Factors; Support, Non-U.S. Gov't; Transformation, Bacterial
Año:1988
Volumen:139
Número:4
Página de inicio:403
Página de fin:409
DOI: http://dx.doi.org/10.1016/0769-2609(88)90103-2
Título revista:Annales de l'Institut Pasteur. Microbiologie
ISSN:07692609
CAS:Culture Media; R Factors
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_07692609_v139_n4_p403_SanchezRivas

Referencias:

  • Chang, Cohen, High frequency transformation of Bacillus subtilis protoplasts by plasmid DNA (1979) Mol. gen. Genetics, 168, pp. 111-115
  • Cofre, Sanchez-Rivas, Curing of plasmids during a protoplast-cell wall regeneration cycle in Bacillus subtilis strains (1983) FEMS Microbiol. Letters, 19, pp. 11-15
  • De Castro-Costa, Landman, Inhibitory protein controls the reversion of protoplasts and L forms of Bacillus subtilis to the walled state (1977) J. Bact., 129, pp. 678-679
  • Duerksen, Localization of the site of fixation of the inducer, penicillin, in Bacillus cereus (1964) Biochim. biophys. Acta (Amst.), 87, pp. 123-140
  • Fowler Edger, Orsech McDonald, Burke, Jr., Plasmid curing in Bacillus subtilis (1981) FEMS Microbiol. Letters, 12, pp. 131-133
  • Gruss, Novick, Plasmid instability in regenerating protoplasts of Staphylococcus aureus is caused by aberrant cell division (1986) J. Bact., 165, pp. 878-883
  • Horinouchi, Weisblum, Nucleotide sequence and functional map of pC194, a plasmid that specifies inducible chloramphenicol resistance (1982) J. Bact., 150, pp. 815-825
  • Hotchkiss, Gabor, Biparental products of bacterial protoplasts fusion showing unequal parental chromosome expression (1980) Proc. nat. Acad. Sci. (Wash), 77, pp. 3553-3557
  • Levi, Sanchez-Rivas, Schaeffer, Further genetic studies on the fusion of bacterial protoplasts (1981) FEMS Microbiology Letters, 2, pp. 323-326
  • Miteva, Shivarova, Grioorova, Transformation of Bacillus thuringiensis protoplasts by plasmid DNA (1981) FEMS Microbiol. Letters, 12, pp. 253-256
  • Rudolph, Schmidt, Saunders, Transformation of Bacillus subtilis by single stranded plasmid DNA (1986) J. Bact., 165, pp. 1015-1018
  • Salermo, Lampen, Transcriptional analysis of β-lactamase regulation in Bacillus licheniformis (1986) J. Bact., 166, pp. 769-778
  • Sanchez-Rivas, Cofré, Symposium on the impact of plasmids on genetics (1979) Lunteren Lectures on Molecular Genetics
  • Sanchez-Rivas, Garro, Bacterial fusion assayed by a prophage complementation test (1979) J Bacteriol, 137, pp. 1340-1345
  • Schaeffer, Cami, Hotchkiss, Fusion of bacterial protoplasts (1976) Proc. nat. Acad. Sci. (Wash), 73, pp. 2151-2155
  • Vos, Venema, Fate of plasmid DNA in transformation of Bacillus subtilis protoplasts (1981) Molecular and General Genetics MGG, 182, pp. 39-43

Citas:

---------- APA ----------
(1988) . Expression and maintenance of plasmid resistance in regenerating protoplasts of Bacillus subtilis. Annales de l'Institut Pasteur. Microbiologie, 139(4), 403-409.
http://dx.doi.org/10.1016/0769-2609(88)90103-2
---------- CHICAGO ----------
Sanchez-Rivas, C. "Expression and maintenance of plasmid resistance in regenerating protoplasts of Bacillus subtilis" . Annales de l'Institut Pasteur. Microbiologie 139, no. 4 (1988) : 403-409.
http://dx.doi.org/10.1016/0769-2609(88)90103-2
---------- MLA ----------
Sanchez-Rivas, C. "Expression and maintenance of plasmid resistance in regenerating protoplasts of Bacillus subtilis" . Annales de l'Institut Pasteur. Microbiologie, vol. 139, no. 4, 1988, pp. 403-409.
http://dx.doi.org/10.1016/0769-2609(88)90103-2
---------- VANCOUVER ----------
Sanchez-Rivas, C. Expression and maintenance of plasmid resistance in regenerating protoplasts of Bacillus subtilis. 1988;139(4):403-409.
http://dx.doi.org/10.1016/0769-2609(88)90103-2