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

Coprinus congregatus did not show any growth on a minimal medium in the pre sence of phosphinothricin which inhibited glutamine synthetase. Genetic tra nsformation to phosphinothricin resistance in C. congregatus was carried ou t successfully by restriction enzyme-mediated integration. The procedure wa s improved to yield 550 transformants per mu g of DNA, and three laccase mu tants were generated. The vector pBARGEM 7-1 which had the phosphinothricin resistance gene was detected in the restriction enzyme fragments of chromo somal DNA from the transformants by Southern hybridization. Transformants s howed identical electrophoretic banding patterns but CL1430b had a faster m oving band when analyzed by native PAGE. (C) 1999 Federation of European Mi crobiological Societies. Published by Elsevier Science B.V. All rights rese rved.