Stability function in the Bacillus subtilis plasmid pTA 1060.
Plasmid pBB2 (11.3 kb) was constructed by genetically labeling the cryptic Bacillus subtilis plasmid pTA 1060 with the pC194-derived CmR and the pUB110-derived KmR markers. In nonselective media pBB2 was segregationally almost completely stable (loss rates less than or equal to 0.02% per cell generation). In contrast, pBB3, obtained by deleting from pBB2 a region consisting of two ClaI fragments (1.45 and 0.20 kb, respectively), was unstable (loss rates greater than or equal to 0.5% per cell generation). This indicates that a genetic element required for stability is located on one or both of these fragments. In pBB3 cop, a mutant with a two- to threefold increased copy number, the rate of plasmid loss was reduced compared to that of pBB3. The insertion of a 4.2-kb Escherichia coli DNA fragment reduced the stability of pBB2 only slightly, suggesting that this vector may be useful for the cloning of relatively large fragments.
Bron
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Department of Genetics, Center of Biological Sciences, Haren, The Netherlands.
Bosma
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van Belkum
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Luxen
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D001412:Bacillus subtilis / Q000235:genetics*; D003001:Cloning, Molecular; D004261:DNA Replication; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000302:isolation & purification; D005838:Genotype; D010957:Plasmids
D004251:DNA Transposable Elements; D004269:DNA, Bacterial