title: A leader open reading frame is essential for the expression in Escherichia coli of GC-rich leuB gene of an extreme thermophile, Thermus thermophilus.
abstract: To improve expression efficiency of the leuB gene of an extreme thermophile. Thermus thermophilus, in Escherichia coli, the gene was placed under a potent promoter, tac. However, the expression was hardly improved, despite increased transcription. The expression under tac promoter was significantly improved by introducing a leader open reading frame in front of the gene. Similar improvement under a weak promoter, tet, with a leader open reading frame had been described previously. The present results provide evidence that the major limiting step in the expression of a GC-rich thermophile gene in E. coli is translation, and that the addition of a leader open reading frame is more crucial for high level expression of the gene than the use of a potent promoter.
authors.affiliation: Laboratory of Biochemistry of Marine Resources, Tokyo University of Fisheries, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Ishida
authors.forename: T
authors.initials: T
authors.lastname: Oshima
mesh_terms: D000943:Antigens, Differentiation / Q000096:biosynthesis / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D016366:Open Reading Frames / Q000502:physiology*; D011401:Promoter Regions, Genetic / Q000502:physiology; D016963:Thermus thermophilus / Q000235:genetics*
chemical_list: D000943:Antigens, Differentiation