title: Selection and characterization of promoters based on genomic approach for the molecular breeding of oleaginous fungus Mortierella alpina 1S-4.
abstract: To express a foreign gene effectively, a good expression system is required. In this study, we investigated various promoters as useful tools for gene manipulation in oleaginous fungus Mortierella alpina 1S-4. We selected and cloned the promoter regions of 28 genes in M. alpina 1S-4 on the basis of expression sequence tag abundance data. The activity of each promoter was evaluated using the β-glucuronidase (GUS) reporter gene. Eight of these promoters were shown to enhance GUS expression more efficiently than a histone promoter, which is conventionally used for the gene manipulation in M. alpina. Especially, the predicted protein 3 and the predicted protein 6 promoters demonstrated approximately fivefold higher activity than the histone promoter. The activity of some promoters changed along with the cultivation phase of M. alpina 1S-4. Seven promoters with constitutive or time-dependent, high-level expression activity were selected, and deletion analysis was carried out to determine the promoter regions required to retain activity. This is the first report of comprehensive promoter analysis based on a genomic approach for M. alpina. The promoters described here will be useful tools for gene manipulation in this strain.
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; D032621:DNA Shuffling; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D015966:Gene Expression Regulation, Fungal; D023061:Gene Order; D017930:Genes, Reporter; D005822:Genetic Vectors; D023281:Genomics; D020094:Mortierella / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D017384:Sequence Deletion; D015533:Transcriptional Activation
chemical_list: D005656:Fungal Proteins