Abstract
Theβ−tubulin gene was isolated from the filamentous fungusAcremonium chrysogenum using a heterologous gene probe to screen anA. chrysogenum lambda library. Sequencing of theA. chrysogenum gene revealed a mosaic gene which contains five exons and four intervening sequences. The exons encode for a polypeptide of 447 amino-acid residues which showed a high degree of similarity when compared with amino-acid sequences from β-tubulins of other eukaryotes. The introns are characterized by typical consensus sequences found in intervening sequences from other filamentous fungi. In-vitro mutagenesis of codon 167 of the β-tubulin gene resulted in the substitution of a phenylalanine by a tyrosine in the corresponding polypeptide sequence. The mutated gene was used successfully in the transformation and co-transformation ofA. chrysogenum to benomyl resistance. The molecular analysis of transformants provided evidence that they contain the mutated β-tubulin gene in addition to the wild-type gene, as was proved by Southern-hybridization analysis and direct sequencing of PCR amplification products.
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Nowak, C., Kück, U. Development of an homologous transformation system forAcremonium chrysogenum based on theβ−tubulin gene. Curr Genet 25, 34–40 (1994). https://doi.org/10.1007/BF00712964
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DOI: https://doi.org/10.1007/BF00712964