Elsevier

Gene

Volume 73, Issue 1, 15 December 1988, Pages 227-235
Gene

The T7 phage gene 10 leader RNA, a ribosome-binding site that dramatically enhances the expression of foreign genes in Escherichia coli

https://doi.org/10.1016/0378-1119(88)90329-0Get rights and content

Abstract

Expression of foreign genes in Escherichia coli requires the juxtaposition of prokaryotic transcription and translation elements with a coding region for the foreign gene. Commonly, this results m only modest expression of the foreign gene product. Here we describe a novel ribosome-binding site (RBS; phage T7 ‘gene 10 leader’) which is able to drive the translation of several foreign genes. This RBS dramatically enhanced the translation efficiency of all the genes we have tested to date, and was particularly effective for foreign genes. The enhanced expression was often more than 40-fold greater than that obtained using a ‘consensus’ RBS. A general plasmid vector has been constructed, incorporating the T7 gene 10 leader sequence, which allows the facile expression of important gene products. In this report we demonstrate the application of this system for the high-level expression of plant, mammalian and bacterial proteins in E. coli.

References (45)

  • T.D. Schneider et al.

    Information content of binding sites on nucleotide sequences

    J. Mol. Biol.

    (1986)
  • J.L. Schottel et al.

    Effects of alterations in the translation initiation control region on bacterial gene expression: use of cat gene constructs transcribed from the lac promoter as a model system

    Gene

    (1984)
  • X. Soberón et al.

    Construction and characterization of new cloning vehicles, IV. Deletion derivatives of pBR322 and pBR325

    Gene

    (1980)
  • P. Stanssens et al.

    Alterations upstream from the Shine-Dalgarno region and their effect on bacterial gene expression

    Gene

    (1985)
  • P. Stanssens et al.

    Inefficient translation initiation causes premature transcription termination in the lacZ gene

    Cell

    (1986)
  • E.A. Whitehorn et al.

    The effects of hybrid ribosome-binding-site variants on the expression of human interferon-β in Escherichia coli

    Gene

    (1985)
  • N.K. Alton et al.

    Nucleotide sequence analysis of the chloramphenicol resistance transposon Tn9

    Nature

    (1979)
  • H.A. de Boer et al.

    Portable Shine-Dalgarno regions: a system for a systematic study of defined alterations of nucleotide sequences within E. coli ribosome binding sites

    DNA

    (1983)
  • G. Buell et al.

    Optimizing the expression in E. coli of a synthetic gene encoding somatomedin-C (IGF-I)

    Nucleic Acids Res.

    (1985)
  • S.I. Feinstein et al.

    Expression of human interferon genes using the recA promoter of E. coli

    Nucleic Acids Res.

    (1983)
  • T.G. Flynn et al.

    The biochemistry and molecular biology of atrial natriuretic factor

    Biochem. J.

    (1985)
  • T.J. Foster et al.

    Chloramphenicol acetyl transferases specified by fi- R-factors

    Antimicrob. Agents Chemother.

    (1973)
  • Cited by (0)

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