, Volume 18, Issue 1, pp 71-76

The development of a heterologous transformation system for the cellulolytic fungus Trichoderma reesei based on a pyrG -negative mutant strain

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Six uridine auxotroph mutants of Trichoderma reesei QM 9414 were isolated by resistance to 5-fluoroorotic acid and one strain was identified as OMP-decarboxylase negative (pyr -) by a radiometric enzyme assay. Transformation to uridine prototrophy was achieved with the pyr4 gene of Neurospora crassa (up to 1500 transformants/μg) and with pyrA of Aspergillus niger (700–800 transformants/μg). In many transformants the PYR+ function seems to be present as extrachromosomal DNA. There is evidence for a correlation between the stability of transformants and integration of the vector in the genome whereas unstable transformants are obtained when autonomous replication of the plasmid occurs.