Electroporation of germinated conidia and young mycelium as an efficient transformation system for Acremonium chrysogenum
Language English Country United States Media print-electronic
Document type Comparative Study, Journal Article
Grant support
CB-2008-01 105527
CONACyT
203440
CONACyT
PubMed
29938299
DOI
10.1007/s12223-018-0625-0
PII: 10.1007/s12223-018-0625-0
Knihovny.cz E-resources
- MeSH
- Acremonium drug effects genetics metabolism MeSH
- Drug Resistance, Bacterial MeSH
- Cephalosporins biosynthesis MeSH
- Electroporation methods MeSH
- Phleomycins pharmacology MeSH
- Microbial Viability MeSH
- Mycelium drug effects genetics metabolism MeSH
- Protoplasts physiology MeSH
- Spores, Fungal drug effects genetics metabolism MeSH
- Transformation, Genetic * MeSH
- Publication type
- Journal Article MeSH
- Comparative Study MeSH
- Names of Substances
- Cephalosporins MeSH
- cephalosporin C MeSH Browser
- Phleomycins MeSH
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.
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