Transformation and laccase mutant isolation in Coprinus congregatus by restriction enzyme-mediated integration

Abstract

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 Wg 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. z

Enhanced saccharification of corn straw pretreated by alkali combining crude ligninolytic enzymes

“…These were in good agreement with other studies. 22,39,40 The colonies germinating from YJ9 protoplasts were then screened by clamp connection observation and DCPI staining to identify monokaryons. 25 Approximately 83 colonies were screened and only 14 were shown to be monokaryotic.…”

Laccases and Their Applications: A Patent Review

“…These included laccases from A. nidulans [99], Coriolus hirsutus [60,62], Phlebia radiata [100], Agaricus bisporus [101], P. cinnabarinus [102], Coriolus versicolor [103], T. versicolor [72], Podospora anserina [104], Coprinus congregatus [105], Ganoderma lucidum, Phlebia brevispora, Lentinula edodes and Lentinus tigrinus Place for Table 2 [ 42,106]. Since then, the number of laccase genes sequenced has increased considerably, and searches from protein and gene sequence databases currently yield several hundreds of laccase gene sequences.…”

Transformation of the medicinal basidiomycete Trametes versicolor to hygromycin B resistance by restriction enzyme mediated integration

“…DNA mediated transformation of basidiomycetes has been reported in Agaricus bisporus [12], Pleurotus ostreatus [13] and Coprinus congregatus [14]. There are many di¡er-ent kinds of selectable markers for transformants, and hygromycin B resistance has been frequently used [12,15].…”

“…During the fungal transformation, the plasmids are usually integrated into the host chromosomes. Many mutants can be obtained during the fungal transformation by restriction enzyme mediated integration (REMI) [14], and part of the mutant gene can be analyzed because it is tagged with the plasmid gene [16]. Therefore, REMI is a very good method to clone genes from various mutants when there are no mutant strains available in an organism.…”