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The Structure and Source of Plasmid DNA Determine the Cloning Properties of Vectors for Bacillus Subtilis

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Book cover Plasmids in Bacteria

Abstract

The elaboration of a system for cloning in bacilli began with a search for plasmids capable of replicating in Bacillus subtilis cells. Various strains of bacilli often carry such plasmids, but most of them are either cryptic or their markers are not selective enough (for review, see Ref. 1,2). Although these plasmids are usable as vectors (2), major progress in gene engineering with bacilli is associated with the use of vectors originating from a phylogenetically different group of organisms. It was found that staphylococcal plasmids can transform B. subtilis cells and render them drug-resistant (3). The plasmids are maintained in a large number of copies in B. subtilis and carry some unique restriction sites suitable for cloning (4,5,6). Several genes were successfully cloned on staphylococcal plasmids as vectors, although as experimentation was extended, it was found that the efficiency of cloning in B. subtilis is much lower than in the traditional Escherichia coli system (4,7,8). Part of the reason for that has become clear after it was demonstrated that subtilis, unlike E. coli, cannot be transformed with plasmid monomers which constitute the bulk of plasmid populations isolated from cells or produced in routine in vitro ligation of DNA fragments. Accordingly, staphylococcal plasmids were found to transform B. subtilis only as oligomers (9,10), although both oligomers and monomers penetrate into the cell equally well.

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Authors and Affiliations

  1. Institute of Genetics and Selection of Industrial Microorganisms, Moscow, 113545, USSR

    P. M. Rabinovich, M. Ya. Haykinson, L. S. Arutyunova, Yu. V. Yomantas & A. I. Stepanov

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  1. P. M. Rabinovich
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  2. M. Ya. Haykinson
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  3. L. S. Arutyunova
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  4. Yu. V. Yomantas
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  5. A. I. Stepanov
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Editor information

Editors and Affiliations

  1. University of California, San Diego, La Jolla, California, USA

    Donald R. Helinski

  2. Stanford University, Stanford, California, USA

    Stanley N. Cohen

  3. University of Michigan, Ann Harbor, Michigan, USA

    Don B. Clewell

  4. Genex Corporation, Gaithersburg, Maryland, USA

    David A. Jackson

  5. Council for Research Planning in Biological Sciences, Inc., Washington, D.C., USA

    Alexander Hollaender

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© 1985 Plenum Press, New York

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Rabinovich, P.M., Haykinson, M.Y., Arutyunova, L.S., Yomantas, Y.V., Stepanov, A.I. (1985). The Structure and Source of Plasmid DNA Determine the Cloning Properties of Vectors for Bacillus Subtilis . In: Helinski, D.R., Cohen, S.N., Clewell, D.B., Jackson, D.A., Hollaender, A. (eds) Plasmids in Bacteria. Springer, Boston, MA. https://doi.org/10.1007/978-1-4613-2447-8_44

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  • DOI: https://doi.org/10.1007/978-1-4613-2447-8_44

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4612-9487-0

  • Online ISBN: 978-1-4613-2447-8

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