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Requirements for the high-level expression of murine interleukin-3 cDNA inEscherichia coli

  • Original Papers
  • Published:
Journal of Industrial Microbiology

Summary

Murine interleukin-3 (Mu IL-3) cDNA was previously expressed inEscherichia coli using atac promoter and a constitutive high copy number plasmid vector. We found that significant increases in expression levels could be realized by using thetac promoter for the expression of Mu IL-3 in a plasmid vector possessing a temperature-inducible runaway-replicon. In contrast, use of anlpp promoter under similar conditions did not result in an increase in the Mu IL-3 expression level. Significant differences were observed when the expression levels of IL-3 were monitored in variousE. coli hosts having different genetic backgrounds. A mutant ofE. coli which lacks the protease La was found to increase the level of IL-3 produced. This report describes the effect of a specific protease-deficientE. coli host strain, as well as the effect of different promoters and plasmid replicons on the expression levels and stability of a heterologous gene product.

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Author notes

  1. Keith M. Gewain

    Present address: Merck, Sharpe & Dohme Research Laboratories, 07065-0900, Rahway, NJ, USA

Authors and Affiliations

  1. Schering Corporation, 60 Orange Street, 07003, Bloomfield, NJ, USA

    Robert Greenberg, Keith M. Gewain, Karen Joy Shaw, Beth Frommer, James A. Anagnost, Satwant K. Narula & Paul J. Leibowitz

  2. DNAX Research Institute of Molecular and Cellular Biology, Inc., Palo Alto, CA, USA

    Sandra Zurawski & Gerard Zurawski

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  1. Robert Greenberg
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Greenberg, R., Gewain, K.M., Shaw, K.J. et al. Requirements for the high-level expression of murine interleukin-3 cDNA inEscherichia coli . Journal of Industrial Microbiology 3, 333–341 (1988). https://doi.org/10.1007/BF01569554

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  • DOI: https://doi.org/10.1007/BF01569554

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