PMID: 8598270Jan 1, 1996Paper

A leader open reading frame is essential for the expression in Escherichia coli of GC-rich leuB gene of an extreme thermophile, Thermus thermophilus

FEMS Microbiology Letters
Masami Ishida, T Oshima

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.

References

Jan 1, 1979·Methods in Enzymology·D A Morrison
Jan 1, 1987·Methods in Enzymology·F Cobianchi, S H Wilson
Jul 25, 1989·Nucleic Acids Research·R A Spanjaard, J van Duin

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Citations

Nov 25, 2006·Extremophiles : Life Under Extreme Conditions·Snaedis H BjornsdottirGudmundur Eggertsson
Aug 15, 2002·Microbiology·Philip J L BellPeter L Bergquist
Sep 3, 1999·Applied and Environmental Microbiology·O FridjonssonR Mattes

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