Agrobacterium-tumefaciens-mediated transformation of antifungal-lipopeptide-producing fungus Coleophoma empetri F-11899.
The filamentous fungus Coleophoma empetri F-11899 produces an echinocandin-like compound FR901379, the original source for micafungin which is prescribed to treat deep-seated mycoses. Despite its industrial importance, no genetic information on C. empetri F-11899 is currently available. To characterize FR901379 biosynthetic genes by insertional mutagenesis and to improve the compound production genetically, Agrobacterium tumefaciens-mediated transformation (ATMT) was attempted to make genetic manipulation possible in this strain. The optimum conditions for ATMT of C. empetri were determined for the cell density of bacteria, time period of co-cultivation and types of filters in co-cultivation. Using the established ATMT method, the hygromycin B resistant gene was successfully transferred into the genome of C. empetri F-11899 and stably maintained even after a serial passage. Some of these results will be applicable for ATMT of various filamentous fungi.
Yamada
Masato
M
Department of Biotechnology, Faculty of Engineering, Toyama Prefectural University, Imizu, Toyama, Japan. myamada@pu-toyama.ac.jp
Yawata
Kazunobu
K
Orino
Yohsuke
Y
Ueda
Satoshi
S
Isogai
Yasuhiro
Y
Taguchi
Goro
G
Shimosaka
Makoto
M
Hashimoto
Seiji
S
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D000935:Antifungal Agents; D004271:DNA, Fungal / Q000235:genetics* / Q000302:isolation & purification; D029968:Escherichia coli Proteins / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics*; D005800:Genes, Fungal; D005822:Genetic Vectors / Q000235:genetics; D006921:Hygromycin B / Q000494:pharmacology; D016254:Mutagenesis, Insertional; D009172:Mycology / Q000379:methods; D010456:Peptides, Cyclic / Q000096:biosynthesis*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics; D004718:Saccharomycetales / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D014170:Transformation, Genetic
D000935:Antifungal Agents; D004271:DNA, Fungal; D029968:Escherichia coli Proteins; C110368:FR 901379; D005656:Fungal Proteins; D010456:Peptides, Cyclic; D006921:Hygromycin B; D017853:Phosphotransferases (Alcohol Group Acceptor); C063242:hygromycin-B kinase