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Detection and characterization of naturally occuring plasmids in Bacillus subtilis

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Summary

Among 83 B. subtilis strains screened, eight were shown to have a significant amount of their DNA in the form of covalently closed circular molecules. One of these strains (1264) carries two such molecular species (plasmids pGY31 and pGY32). A group of four strains (14A1, X35, X36, X43) each carry one plasmid (pGY5 to pGY8). These four plasmids were found to be very similar to each other. Strain 2588 carries a large plasmid (pGY1). The satellite DNA's of the remaining two strains (11A1 and 13F3) are very heterogeneous. The plasmids were isolated and characterized as regards sedimentation velocity, copy number per chromosome equivalent and EcoRI cleavage pattern. The sedimentation coefficient of pGY1 suggested a molecular weight of the order of 40 Md. This plasmid is present in no more than two copies per chromosome. It possesses many EcoRI sites. Plasmids pGY5 to pGY8 each have two EcoRI sites. Cleavage pattern and sedimentation coefficient are the same in all four of them. Their estimated M.W. is 4.9 Md. pGY7 of this class was studied in more detail. Its average copy number is 20. pGY31 and pGY32 both have a single EcoRI site. Their M.W.'s are respectively 7.6 and 3.6 Md and the minimal copy numbers are 7 and 15. Contour lengths were determined for pGY7 (=2.94 μ), pGY31 (=4.46 μ) and pGY32 (=1.84 μ). No phenotype associated with any of the above plasmids has so far been detected. Among the eight strains harboring plasmids only one (2588, plasmid pGY1) was found to be genetically homologous to strain 168, by transformation. Strains 1264 and X36 are indistinguishable from 168 in physiological tests.

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Communicated by W. Arber

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Le Hégarat, J., Anagnostopoulos, C. Detection and characterization of naturally occuring plasmids in Bacillus subtilis . Molec. Gen. Genet. 157, 167–174 (1977). https://doi.org/10.1007/BF00267394

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Keywords

  • Bacillus
  • Bacillus Subtilis
  • Sedimentation Velocity
  • EcoRI Site
  • Contour Length