An improved transformation vector for the lignin-degrading white rot basidiomycete Phanerochaete chrysosporium.
In this study, a lignin peroxidase-encoding gene (LIP) of Phanerochaete chrysosporium was disrupted by inserting into its coding region the kanamycin-resistance determinant from Tn903. The resulting recombinant plasmid, pUGLG1: kan, was transformed into P. chrysosporium with the expectation that the disrupted gene might replace the homologous LIP gene in the chromosome. However, the results showed that pUGLG1: kan sequences do not integrate into the chromosome; instead, the plasmid is maintained intact in the transformants in an extrachromosomal state. Our data also show that pUGLG1: kan undergoes replication in P. chrysosporium, is maintained as a circular element, is recoverable from meiotic and mitotic progeny, although at a low frequency, and can be recovered intact by Escherichia coli transformation. These results suggest that the GLG1 component of pUGLG1: kan contains as yet unidentified sequences that allow its autonomous replication in P. chrysosporium transformants.
Randall
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T
Department of Microbiology and Public Health, Michigan State University, East Lansing 48824-1101.
Reddy
C A
CA
D001487:Basidiomycota / Q000235:genetics* / Q000378:metabolism; D015139:Blotting, Southern; D004926:Escherichia coli / Q000235:genetics; D005800:Genes, Fungal / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics*; D007613:Kanamycin Resistance / Q000235:genetics; D008031:Lignin / Q000378:metabolism; D010544:Peroxidases / Q000235:genetics*; D010957:Plasmids / Q000235:genetics*; D014170:Transformation, Genetic / Q000235:genetics*
D008031:Lignin; D010544:Peroxidases; C042858:lignin peroxidase