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Molecular and General Genetics MGG

, Volume 217, Issue 2–3, pp 488–498 | Cite as

Characterization of the ColE1 mobilization region and its protein products

  • A. Christopher Boyd
  • John A. K. Archer
  • David J. Sherratt
Article

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.

Key words

ColE1 Plasmid mobilization Overlapping genes Site-directed mutagenesis Relaxation complex 

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Copyright information

© Springer-Verlag 1989

Authors and Affiliations

  • A. Christopher Boyd
    • 1
  • John A. K. Archer
    • 2
  • David J. Sherratt
    • 3
  1. 1.Department of Molecular BiologyUniversity of EdinburghEdinburghUK
  2. 2.Department of Applied Biological SciencesMassachusetts Institute of TechnologyCambridgeUSA
  3. 3.Institute of GeneticsUniversity of GlasgowGlasgowUK

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