Characterization of galactose-dependent promoters from an oleaginous fungus Mortierella alpina 1S-4

被引：7

作者：

Okuda, Tomoyo ^{[1
]}

Ando, Akinori ^{[1
,2
]}

Sakuradani, Eiji ^{[1
]}

Kikukawa, Hiroshi ^{[1
]}

Kamada, Nozomu ^{[3
]}

Ochiai, Misa ^{[3
]}

Shima, Jun ^{[4
]}

Ogawa, Jun ^{[1
]}

机构：

[1] Kyoto Univ, Grad Sch Agr, Div Appl Life Sci, Sakyo Ku, Kyoto 6068052, Japan

[2] Kyoto Univ, Res Unit Physiol Chem, Sakyo Ku, Kyoto 6068052, Japan

[3] Suntory Global Innovat Ctr Ltd, Res Inst, Shimamoto, Osaka 6188503, Japan

[4] Kyoto Univ, Res Div Microbial Sci, Sakyo Ku, Kyoto 6068052, Japan

来源：

CURRENT GENETICS | 2014年 / 60卷 / 03期

关键词：

Mortierella alpina; Inducible promoter; Galactose; Polyunsaturated fatty acid; Zygomycete; HETEROLOGOUS PROTEIN-PRODUCTION; POLYUNSATURATED FATTY-ACIDS; REGULATED EXPRESSION; FILAMENTOUS FUNGI; DNA-BINDING; GENE; TRANSFORMATION; OIL; DESATURASE; GLUCOSE;

D O I：

10.1007/s00294-014-0422-5

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

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 beta-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.

引用

页码：175 / 182

页数：8

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