Genetic transformation and mutant isolation in Ganoderma lucidum by restriction enzyme-mediated integration

Abstract A white-rot basidiomycete Ganoderma lucidum has long been used as a medicinal mushroom in Asia, and it has an array of enzymes important for wood degrading activity. There have been many reports about the ingredients which show health aiding effects. In order to analyze gene functions and i...

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Bibliographic Details
Published in:FEMS microbiology letters 2004-04, Vol.233 (2), p.201-204
Main Authors: Kim, Sunkyung, Song, Jaemahn, Choi, Hyoung T.
Format: Article
Language:English
Subjects:
Antibiotic resistance
Antibiotics
Basidiocarps
Biological and medical sciences
Chromosomes
Cloning, Molecular
Deoxyribonuclease EcoRI - genetics
Deoxyribonucleases, Type II Site-Specific - genetics
Enzymes
Fundamental and applied biological sciences. Psychology
Fungal transformation
Fungi
Ganoderma lucidum
Genes
Genetic Testing - methods
Genetic transformation
Geneticin
Growth, nutrition, metabolism, transports, enzymes. Molecular biology
Integration
Microbiology
Miscellaneous
Mushrooms
Mutation
Mycology
Phosphinothricin
Plasmids
Reishi - genetics
Rot
Transformation, Genetic
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Summary:Abstract A white-rot basidiomycete Ganoderma lucidum has long been used as a medicinal mushroom in Asia, and it has an array of enzymes important for wood degrading activity. There have been many reports about the ingredients which show health aiding effects. In order to analyze gene functions and introduce foreign genes into this fungus, genetic transformation is required. We have successfully transformed G. lucidum to geneticin resistance using pJS205-1 which has the antibiotic resistance genes against geneticin and phosphinothricin. Many different mutants have been generated during the transformation by restriction enzyme mediated integration, and the transformation yield was 4–17 transformants (μg plasmid DNA)−1. The plasmid was integrated stably into the recipient chromosome, which was confirmed by PCR with the plasmid-specific primers.
ISSN:0378-1097
1574-6968
DOI:10.1111/j.1574-6968.2004.tb09483.x