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Characterization of the ColE1 mobilization region and its protein products

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Summary

A third of the 6.6 kb genome of ColE1 is devoted to mobilization (mob) genes necessary to promote its specific transfer in the presence of conjugative plasmids. Themob region is genetically complex: twomob genes are entirely overlapped by a third. Oligonucleotide-directed mutagenesis was used to insert an amber codon into one of the overlapped genes and make possible a full complementation analysis ofmob. Fourmob genes essential for mobilization by R64drd11 were thus identified. Fragments ofmob were subcloned under control of the Ptac promoter in a suitable vector, overexpressed in minicells and the mobilization proteins visualized. A comprehensive alignment of themob region of ColE1 with those of its close relatives ColK and ColA demonstrating that the four essentialmob genes are conserved is also presented.

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Authors and Affiliations

  1. Department of Molecular Biology, University of Edinburgh, King’s Buildings, Mayfield Rd., EH9 3JR, Edinburgh, UK

    A. Christopher Boyd

  2. Department of Applied Biological Sciences, Massachusetts Institute of Technology, Room 56-121, 02139, Cambridge, MA, USA

    John A. K. Archer

  3. Institute of Genetics, University of Glasgow, Church St., G111 5JS, Glasgow, UK

    David J. Sherratt

Authors
  1. A. Christopher Boyd
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  2. John A. K. Archer
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  3. David J. Sherratt
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Communicated by W. Goebel

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Christopher Boyd, A., Archer, J.A.K. & Sherratt, D.J. Characterization of the ColE1 mobilization region and its protein products. Molec. Gen. Genet. 217, 488–498 (1989). https://doi.org/10.1007/BF02464922

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  • DOI: https://doi.org/10.1007/BF02464922

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