A leader open reading frame is essential for the expression in Escherichia coli of GC-rich leuB gene of an extreme thermophile, Thermus thermophilus.
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.
Ishida
M
M
Laboratory of Biochemistry of Marine Resources, Tokyo University of Fisheries, Japan.
Oshima
T
T
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*
D000943:Antigens, Differentiation