Summary
The Trichoderma reesei orotidine-5′-phosphate decarboxylase gene was isolated by heterologous hybridization with the corresponding Neurospora gene as a probe. A 2.7 kb SalI fragment, which exclusively hybridized to the Neurospora gene, was subcloned in pGEM-5Zf(+). This subclone was termed pFG1 and was used to transform a Trichoderma reesei pyrG- negative mutant to PYR+. The transformation frequency in this homologous system was up to 12000 transformants per μg DNA. About one-fifth of the transformants tested were abortive. Perfect mitotic stability was found in half of the non-abortive transformants, correlating with vector integration at homologous and ectopic loci. In the unstable transformants the transforming DNA appears to be present in the form of extrachromosomal elements.
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Communicated by R. J. Schweyen
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Gruber, F., Visser, J., Kubicek, C.P. et al. Cloning of the Trichoderma reesei pyrG gene and its use as a homologous marker for a high-frequency transformation system. Curr Genet 18, 447–451 (1990). https://doi.org/10.1007/BF00309915
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DOI: https://doi.org/10.1007/BF00309915