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Construction of expression vectors for the gram-negative bacterium Zymomonas mobilis

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Summary

A set of vectors was constructed for the cloning and expression of heterologous genes in the Gramnegative bacterium Zymomonas mobilis under the control of the pdc promoter of Z. mobilis. The vectors pPTZ1, pPTZ3, and pPTZ4 are based on the cryptic Z. mobilis plasmid pZM02 and on parts of the Escherichia coli plasmids pKK223-3 and pBR322 together with the multiple cloning site of phage Ml3mp18. DNA fragments can be readily inserted immediately downstream from the pdc promoter at unique restriction sites for KpnI, XbaI and PstI in pPTZl and additionally for SmaI and BamHI in pPTZ3. In pPTZ4, the 5′ terminal codons of pdc were deleted allowing the formation of gene fusions. Expression of a promoterless chloramphenicol acetyltransferase gene (cat) controlled by the pdc gene promoter resulted in enzyme activities of up to 5.5 U/mg total cell protein in Z. mobilis cells.

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Authors and Affiliations

  1. Institut für Biotechnologie 1, Forschungszentrum Jülich GmbH, Postfach 1913, D-5170, Jülich, Germany

    Michael Reynen, Ina Reipen, Hermann Sahm & Georg A. Sprenger

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  1. Michael Reynen
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  2. Ina Reipen
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  3. Hermann Sahm
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  4. Georg A. Sprenger
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Communicated by P. Tiollais

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Reynen, M., Reipen, I., Sahm, H. et al. Construction of expression vectors for the gram-negative bacterium Zymomonas mobilis . Molec. Gen. Genet. 223, 335–341 (1990). https://doi.org/10.1007/BF00265073

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  • DOI: https://doi.org/10.1007/BF00265073

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