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Genetic mapping by means of protoplast fusion in Bacillus subtilis

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Summary

A new mapping method involving protoplast fusion in Bacillus subtilis is described. Protoplasts from an isogenic standard marker strain containing purA and from a strain containing both purB and the marker, “x”, to be mapped were fused with polyethylene glycol, and purA + purB + fusants were selected. After isolation of single colonies and determination of unselected markers, marker x was mapped between two standard markers. This method was fully applicable to PBS1-resistant strains (e.g., lyt strains). The results obtained by protoplast fusion, conventional transformation and/or lysed protoplast transformation indicated that a lyt strain, Ni15, contained two new autolysin-minus mutations (lyt-151 and lyt-152). The properties of lyt-15 are also discussed.

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Abbreviations

NTG:

N-methyl-N′-nitro-N-nitrosoguanidine

SMM:

0.5 M sucrose, 0.02 M MgCl2, 0.02 M maleate buffer, pH 6.5

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Authors and Affiliations

  1. Department of Applied Microbiology and Technology, Kumamoto Institute of Technology, Ikeda 4-22-1, 860, Kumamoto, Japan

    T. Akamatsu

  2. Department of Applied Biology, Faculty of Textile Science and Technology, Shinshu University, 3-15-1 Tokida, 386, Ueda, Japan

    J. Sekiguchi

Authors
  1. T. Akamatsu
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  2. J. Sekiguchi
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Communicated by H. Böhme

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Akamatsu, T., Sekiguchi, J. Genetic mapping by means of protoplast fusion in Bacillus subtilis . Mole Gen Genet 208, 254–262 (1987). https://doi.org/10.1007/BF00330451

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  • DOI: https://doi.org/10.1007/BF00330451

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