Open AccessTransformation and laccase mutant isolation in Coprinus congregatus by restriction enzyme‐mediated integration
Author(s) -
Leem Youngeun,
Kim Soonja,
Ross Ian K.,
Choi Hyoung T.
Publication year - 1999
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.1574-6968.1999.tb13446.x
Subject(s) - transformation (genetics) , restriction enzyme , mutant , biology , glutamine synthetase , enzyme , coprinus , microbiology and biotechnology , biochemistry , restriction map , dna , gene , genetics , glutamine , plasmid , botany , amino acid
Coprinus congregatus did not show any growth on a minimal medium in the presence of phosphinothricin which inhibited glutamine synthetase. Genetic transformation to phosphinothricin resistance in C. congregatus was carried out successfully by restriction enzyme‐mediated integration. The procedure was improved to yield 550 transformants per μg of DNA, and three laccase mutants were generated. The vector pBARGEM 7‐1 which had the phosphinothricin resistance gene was detected in the restriction enzyme fragments of chromosomal DNA from the transformants by Southern hybridization. Transformants showed identical electrophoretic banding patterns but CL1430b had a faster moving band when analyzed by native PAGE.