Genetic transformation and mutant isolation in Ganoderma lucidum by restriction enzyme-mediated integration.
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 (microg plasmid DNA)(-1). The plasmid was integrated stably into the recipient chromosome, which was confirmed by PCR with the plasmid-specific primers.
Kim
Sunkyung
S
Microbial Physiology Laboratory, Division of Life Sciences, Kangwon National University, Chunchon 200-701, South Korea.
Song
Jaemahn
J
Choi
Hyoung T
HT
D003001:Cloning, Molecular; D015246:Deoxyribonuclease EcoRI / Q000235:genetics; D015252:Deoxyribonucleases, Type II Site-Specific / Q000235:genetics; D005820:Genetic Testing / Q000379:methods; D009154:Mutation; D010957:Plasmids; D020940:Reishi / Q000235:genetics*; D014170:Transformation, Genetic
D015246:Deoxyribonuclease EcoRI; C025630:CAGCTG-specific type II deoxyribonucleases; C025651:CCCGGG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific