Open AccessGenetic transformation and mutant isolation in Ganoderma lucidum by restriction enzyme‐mediated integration
Author(s) -
Kim Sunkyung,
Song Jaemahn,
Choi Hyoung T.
Publication year - 2004
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2004.tb09483.x
Subject(s) - transformation (genetics) , plasmid , mutant , biology , restriction enzyme , gene , genetics , microbiology and biotechnology
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.