Summary
The thermostability of the staphylococcal plasmids pC194 and pUB110 and their antibiotic-resistance determinants was examined upon transfer to Bacillus stearothermophilus CU21. Plasmid pGS13, a pUB110 derivative carrying the chloramphenicol acetyltransferase (CAT) gene of pC194, could be maintained up to the maximum growth temperature (68° C) by selection for chloramphenicol resistance. In the absence of selective pressure, pGS13 was lost at temperatures above 60° C. Segregational instability of pGS13 was accompanied by a progressive loss of negative superhelicity at elevated temperatures. Thermostable mutants of pGS13 were isolated by screening for expression of the antibiotic-resistance determinants after growth under non-selective conditions. These mutants were found to contain an insertion of a 1.7 kb DNA sequence derived from the cryptic B. stearothermophilus plasmid pBS02. Increased thermostability correlated with preservation of plasmid superhelicity at elevated temperatures.
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Communicated by W. Goebel
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Soutschek-Bauer, E., Scholz, W., Grill, E. et al. Thermostability and superhelicity of plasmid DNA in Bacillus stearothermophilus . Mol Gen Genet 209, 575–579 (1987). https://doi.org/10.1007/BF00331165
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DOI: https://doi.org/10.1007/BF00331165