Use of a linear plasmid containing telomeres as an efficient vector for direct cloning in the filamentous fungus Podospora anserina.
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 integrative vectors. Genomic DNA was inserted in this plasmid in vitro and the mixture used to transform a leu1-1 strain. Many fungal clones containing the leu1 gene as a genomic insert in the linear plasmid were identified. 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.
Barreau
C
C
Laboratoire de Génétique et Biologie Moléculaire des Champignons, Institut de Biochimie et Génétique Cellulaires-UPR CNRS-9026, 1, Rue Camille Saint-Saëns, Bordeaux cedex, 33077, France.
Iskandar
M
M
Turcq
B
B
Javerzat
J P
JP
D001203:Ascomycota / Q000235:genetics* / Q000254:growth & development; D015139:Blotting, Southern; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007930:Leucine / Q000235:genetics; D010957:Plasmids / Q000235:genetics*; D015183:Restriction Mapping; D016615:Telomere / Q000235:genetics*; D014170:Transformation, Genetic
D007930:Leucine