Electroporation of germinated conidia and young mycelium as an efficient transformation system for Acremonium chrysogenum.


Journal

Folia microbiologica
ISSN: 1874-9356
Titre abrégé: Folia Microbiol (Praha)
Pays: United States
ID NLM: 0376757

Informations de publication

Date de publication:
Jan 2019
Historique:
received: 10 05 2017
accepted: 11 06 2018
pubmed: 26 6 2018
medline: 17 5 2019
entrez: 26 6 2018
Statut: ppublish

Résumé

Three different transformation strategies were tested and compared in an attempt to facilitate and improve the genetic transformation of Acremonium chrysogenum, the exclusive producer of the pharmaceutically relevant β-lactam antibiotic cephalosporin C. We investigated the use of high-voltage electric pulse to transform germinated conidia and young mycelium and compared these procedures with traditional PEG-mediated protoplast transformation, using phleomycin resistance as selection marker in all cases. The effect of the field strength and capacitance on transformation frequency and cell viability was evaluated. The electroporation of germinated conidia and young mycelium was found to be appropriate for transforming A. chrysogenum with higher transformation efficiencies than those obtained with the conventional protoplast-based transformation procedures. The developed electroporation strategy is fast, simple to perform, and highly reproducible and avoids the use of chemicals toxic to cells. Electroporation of young mycelium represents an alternative method for transformation of fungal strains with reduced or no sporulation, as often occurs in laboratory-developed strains in the search for high-yielding mutants for industrial bioprocesses.

Identifiants

pubmed: 29938299
doi: 10.1007/s12223-018-0625-0
pii: 10.1007/s12223-018-0625-0
doi:

Substances chimiques

Cephalosporins 0
Phleomycins 0
cephalosporin C 3XIY7HJT5L

Types de publication

Comparative Study Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

33-39

Subventions

Organisme : CONACyT
ID : CB-2008-01 105527
Organisme : CONACyT
ID : 203440

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Auteurs

Jessica Cruz-Ramón (J)

Departamento de Biotecnología, Universidad Autónoma Metropolitana-Unidad Iztapalapa, Ciudad de México, Mexico.

Francisco J Fernández (FJ)

Departamento de Biotecnología, Universidad Autónoma Metropolitana-Unidad Iztapalapa, Ciudad de México, Mexico.

Armando Mejía (A)

Departamento de Biotecnología, Universidad Autónoma Metropolitana-Unidad Iztapalapa, Ciudad de México, Mexico.

Francisco Fierro (F)

Departamento de Biotecnología, Universidad Autónoma Metropolitana-Unidad Iztapalapa, Ciudad de México, Mexico. fierrof@xanum.uam.mx.

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Classifications MeSH