Transformation of Penicillium freii and a rapid PCR screening procedure for cotransformation events.
A strain of Penicillium freii was transformed to hygromycin B resistance using the pAN7-1 plasmid. The transformation frequency was approximately 100 transformants per microgram DNA per 1 x 10(7) protoplasts. Southern blotting revealed different types of integration into the genome of P.freii. When non-selectable plasmid DNA was added together with pAN7-1, this resulted in a cotransformation frequency of more than 60%. The cotransformation frequency was estimated using a facile DNA extraction method together with PCR.
Nicolaisen
M
M
Dept. of Biotechnology, Technical University of Denmark, Lyngby, Denmark.
Geisen
R
R
D015139:Blotting, Southern; D004271:DNA, Fungal / Q000032:analysis; D004352:Drug Resistance, Microbial; D006921:Hygromycin B / Q000378:metabolism; D010407:Penicillium / Q000235:genetics*; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D014169:Transformation, Bacterial
D004271:DNA, Fungal; D006921:Hygromycin B