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Expression of heterologous genes in Schizophyllum commune is often hampered by the formation of truncated transcripts

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Author: Schuren, F.H.J. · Wessels, J.G.H.
Institution: Centraal Instituut voor Voedingsonderzoek TNO
Source:Current Genetics, 2, 33, 151-156
Identifier: 234363
doi: DOI:10.1007/s002940050321
Keywords: Nutrition · Basidiomycete fungus · Heterologous gene expression · Schizophyllum commune · Transcription termination · Aminoglycoside · Beta glucuronidase · Hygromycin b · Messenger rna · Phleomycin · Phosphotransferase · Antibiotic resistance · Fungal genetics · Molecular cloning · Nonhuman · Priority journal · Schizophyllum commune · Base Sequence · beta-Galactosidase · DNA, Fungal · Gene Expression Regulation, Enzymologic · Gene Expression Regulation, Fungal · Glucuronidase · Glyceraldehyde-3-Phosphate Dehydrogenases · Kanamycin Kinase · Molecular Sequence Data · Phosphotransferases (Alcohol Group Acceptor) · Schizophyllum · Fungi · Schizophyllum commune


GPD regulatory sequences were used to express a phleomycin resistance gene (Sh ble) in Schizophyllum commune, resulting in high numbers of phleomycin-resistant transformants. Attempts to express heterologous genes coding for hygromycin B phosphotransferase (hph), aminoglycoside phosphotransferase (apt), β-glucuronidase (uidA) and β-galactosidase (lacZ) using the same regulatory sequences were not successful and no mRNA could be detected. Cloning the hph and uidA genes in an internally deleted GPD gene resulted in truncated transcripts which ended within the 5'-parts of the heterologous genes. Cloning of the same genes as transcriptional fusions downstream from the Sh ble gene also resulted in truncated transcripts ending in the 5'-parts of these heterologous genes. It is suggested that AT-rich sequences in heterologous genes might be involved in generating these truncated transcripts, thereby preventing expression in S. commune.