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Distance-dependent translational coupling and interference inLactococcus lactis

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Summary

The possibility of raising the expression level of a heterologous gene inLactococcus lactis by exploiting the principle of translational coupling was investigated. For this purpose, theEscherichia coli lacZ gene was transcriptionally fused to a short open reading frame (ORF) of lactococcal origin. A Shine-Dalgarno (SD) sequence was introduced at the boundary of the two ORFs. In a series of otherwise identical plasmids, the relative positions of the translational stop codon of the upstream ORF and the translational start codon of the downstream ORF (lacZ) were varied. The expression oflacZ gradually increased as the stop and start codons were placed in closer proximity. A concomitant switch from translational interference to translational coupling was observed. Best results were obtained with partially overlapping stop and start codons. It is concluded that the principle of translational coupling offers good possibilities to increase the level of heterologous gene expression inL. lactis.

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Authors and Affiliations

  1. Department of Genetics, Centre of Biological Sciences, University of Groningen, Kerklaan 30, NL-9751, NN Haren, The Netherlands

    Maarten van de Guchte, Jan Kok & Gerard Venema

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  1. Maarten van de Guchte
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  2. Jan Kok
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  3. Gerard Venema
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Communicated by H. Böhme

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van de Guchte, M., Kok, J. & Venema, G. Distance-dependent translational coupling and interference inLactococcus lactis . Molec. Gen. Genet. 227, 65–71 (1991). https://doi.org/10.1007/BF00260708

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  • DOI: https://doi.org/10.1007/BF00260708

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