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The development of a heterologous transformation system for the cellulolytic fungus Trichoderma reesei based on a pyrG-negative mutant strain
July 1990
July 1990
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Abstract
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/micrograms) and
with pyrA of Aspergillus niger (700-800 transformants/micrograms). 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.
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Most cited references11
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A positive selection for mutants lacking orotidine-5′-phosphate decarboxylase activity in yeast: 5-fluoro-orotic acid resistance
Jef D. Boeke, Francois La Croute, Gerald Fink (1984)
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A versatile transformation system for the cellulolytic filamentous fungus Trichoderma reesei
Merja Penttilä, Jonathan Knowles, Helena Nevalainen … (1987)