A truncated derivative of nmt 1 promoter exhibits temperature-dependent induction of gene expression in Schizosaccharomyces pombe.
Despite increasing exploitation of Schizosaccharomyces pombe as a model system there is a lack of convenient vectors for research and application. Expression with the commonly used promoter, nmt 1, requires a laborious regime involving the removal of repressor, thiamine, from a growing culture and further growth for 18 h to achieve maximum expression, thus underlining the need for more user-friendly promoters. We report here the isolation and characterization of a truncated derivative of the nmt 1 promoter having novel induction characteristics: it is induced by shift of growth temperature from 36 degrees C to 25 degrees C, achieving maximum expression within 3 h. Similar features of expression were observed with the reporter genes GFP and beta-galactosidase, a native gene, cdc 18, and a commercially important foreign therapeutic protein, streptokinase. The new promoter element offers additional advantages, such as lack of deleterious effect on cell viability and potential ability to express toxic proteins. These features make the new promoter a potentially better alternative to nmt 1, both as a research tool and for expression of commercially important proteins in Sz. pombe, and suggest the possibility of using similar approaches to design promoters with novel and useful properties.
Kumar
Raj
R
Institute of Microbial Technology, Sector 39A, Chandigarh-160036, India.
Singh
Jagmohan
J
D001483:Base Sequence; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D005434:Flow Cytometry; D015966:Gene Expression Regulation, Fungal / Q000502:physiology*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000502:physiology*; D012568:Schizosaccharomyces / Q000235:genetics* / Q000378:metabolism; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics* / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013696:Temperature; D014170:Transformation, Genetic / Q000502:physiology
D004271:DNA, Fungal; C093953:NMT1 protein, S pombe; D029702:Schizosaccharomyces pombe Proteins; D049452:Green Fluorescent Proteins