Open AccessA leader open reading frame is essential for the expression in Escherichia coli of GC‐rich leuB gene of an extreme thermophile, Thermus thermophilus
Author(s) -
Ishida Masami,
Oshima Tairo
Publication year - 1996
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1996.tb07978.x
Subject(s) - thermus thermophilus , open reading frame , biology , escherichia coli , gene , thermophile , thermus , gene expression , genetics , microbiology and biotechnology , promoter , transcription (linguistics) , bacteria , peptide sequence , linguistics , philosophy
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.