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An E. coli-yeast shuttle cosmid with positive selection for inserted fragments

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Summary

We describe the construction of a cosmid cloning vector, pMT555, which allows positive selection for the presence of an inserted DNA fragment. The vector contains sequences which enable its replication and selection in either E. coli or Saccharomyces cerevisiae. We demonstrate that pMT555 may be used for the efficient construction of total genomic banks from small quantities of donor DNA. The positive selection permits the stable maintenance of the cosmid in E. coli and the faithful replication of inserted sequences.

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Author notes

  1. J. Richard Ludwig

    Present address: Marine Biology Research Division A-002, Scripps Institution of Oceanography, UCSD, 92093, La Jolla, CA, USA

  2. David A. Sugden

    Present address: Pfizer Central Research, CT13 9NJ, Sandwich, Kent, UK

Authors and Affiliations

  1. Department of Biochemistry and Applied Molecular Biology, University of Manchester Institute of Science and Technology, P. O. Box 88, M60 1QD, Manchester, UK

    Manda E. Gent, Patrick Crowley, J. Richard Ludwig, Rashida Anwar, David A. Sugden, Paul F. G. Sims & Stephen G. Oliver

Authors
  1. Manda E. Gent
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  2. Patrick Crowley
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  3. J. Richard Ludwig
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  4. Rashida Anwar
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  5. David A. Sugden
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  6. Paul F. G. Sims
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  7. Stephen G. Oliver
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Gent, M.E., Crowley, P., Ludwig, J.R. et al. An E. coli-yeast shuttle cosmid with positive selection for inserted fragments. Curr Genet 10, 29–33 (1985). https://doi.org/10.1007/BF00418490

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  • DOI: https://doi.org/10.1007/BF00418490

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