USE OF A LINEAR PLASMID CONTAINING TELOMERES AS AN EFFICIENT VECTOR FOR DIRECT CLONING IN THE FILAMENTOUS FUNGUS PODOSPORA-ANSERINA

Use of linear plasmid containing telomeres as an efficient vector for direct cloning in the filamentous fungus Podospora anserina, Fungal Ge netics and Biology (25, 22-30. In Podospora anserina a linear plasmid with telomeric ends behaves as an artificial acentric minichromosome, Transformation is at least 100 times more efficient than with integrat ive vectors. Genomic DNA was inserted in this plasmid in vitro and the mixture used to transform a leu1-1 strain. Many fungal clones contain ing the leu1 gene as a genomic insert in the linear plasmid were ident ified. The leu1 gene was rescued as a circular plasmid in Escherichia coli demonstrating that a direct cloning procedure can be applied for the fungus P. anserina. The conservation of telomeric sequences among filamentous fungi suggests that a telomere-based linear plasmid could provide a general cloning vector for filamentous fungi. (C) 1998 Acade mic Press.