title: A method for the re-isolation of an autonomously replicating plasmid from Aspergillus transformants.
abstract: An autonomously replicating plasmid is expected to increase the frequency of Aspergillus transformation. To construct this type of plasmid, we developed a rapid method of re-isolating autonomously replicating plasmids from Aspergillus transformants. Transformants grown in MM medium under selective pressure for 1-2 days were converted to protoplasts with a cell wall lytic enzyme (e.g. Yatalase). The protoplasts were lysed with phenol/chloroform followed by precipitation with ethanol. The total DNA was treated with RNaseA, re-precipitated with PEG, and then used to transform E. coli. These re-isolated plasmids were mainly the plasmid monomer.
authors.affiliation: General Research Laboratory, Ozeki Corporation, Hyogo, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Ozeki
authors.forename: K
authors.initials: K
authors.lastname: Hizume
authors.forename: A
authors.initials: A
authors.lastname: Kanda
authors.forename: M
authors.initials: M
authors.lastname: Hamachi
authors.forename: Y
authors.initials: Y
authors.lastname: Nunokawa
mesh_terms: D001230:Aspergillus / Q000235:genetics*; D004261:DNA Replication; D004274:DNA, Recombinant / Q000096:biosynthesis; D010957:Plasmids
chemical_list: D004274:DNA, Recombinant