title: Characterization of galactose-dependent promoters from an oleaginous fungus Mortierella alpina 1S-4.
abstract: An inducible promoter is a useful tool for the controlled expression of a given gene. In this report, we describe galactose-dependent promoters for potential use in an oleaginous fungus Mortierella alpina. We cloned the putative promoter regions of two genes encoding galactose metabolic enzymes, GAL1 and GAL10, from the genome of M. alpina 1S-4. The β-glucuronidase (GUS) reporter gene assay in M. alpina 1S-4 revealed that regulation of these promoters was dependent on the presence of galactose in the medium both with and without other sugars. With the GAL10 promoter, an approximately 50-fold increase of GUS activity was demonstrated by addition of galactose into the culture media at any cultivation phase. The 5' deletion analysis of the GAL10 promoter revealed that a promoter region of over 2,000 bp length was required for its high-level activity and sufficient inducible response. Significantly, this is the first report of inducible promoters of zygomycetes. The GAL10 promoter will be a valuable tool for gene manipulation in M. alpina 1S-4.
authors.affiliation: Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa-oiwakecho, Sakyo-ku, Kyoto, 606-8052, Japan.
authors.forename: Tomoyo
authors.initials: T
authors.lastname: Okuda
authors.forename: Akinori
authors.initials: A
authors.lastname: Ando
authors.forename: Eiji
authors.initials: E
authors.lastname: Sakuradani
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kikukawa
authors.forename: Nozomu
authors.initials: N
authors.lastname: Kamada
authors.forename: Misa
authors.initials: M
authors.lastname: Ochiai
authors.forename: Jun
authors.initials: J
authors.lastname: Shima
authors.forename: Jun
authors.initials: J
authors.lastname: Ogawa
mesh_terms: D003001:Cloning, Molecular; D003470:Culture Media; D005656:Fungal Proteins / Q000235:genetics; D005690:Galactose / Q000378:metabolism*; D015870:Gene Expression; D015966:Gene Expression Regulation, Fungal; D017930:Genes, Reporter; D020094:Mortierella / Q000235:genetics* / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D017384:Sequence Deletion
chemical_list: D003470:Culture Media; D005656:Fungal Proteins; D005690:Galactose