Cloning of sporulation gene spoIIG in Bacillus subtilis

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Abstract
Two specialized transducing phages carrying a sporulation gene, spoIIG, of B. subtilis were constructed from B. subtilis temperate phages .rho.11 and .vphi.105 by the prophage transformation method. Restriction enzyme analysis and transformation experiments showed that the spoIIG gene was present on a 6.2 megadalton (Md) EcoRi fragment in both transducing phage genomes. Further analysis showed that spoIIG+ transforming activity resides on a 2.25-Md EcoRI-BamHI fragment within the 6.2-Md EcoRI fragment. The 2.25-Md fragment was subcloned into the region between the EcoRI and BamHI sites of pUB110, and deletion plasmids lacking PstI or HindIII fragments within the 2.25-Md fragment were constructed. The recombinant plasmid carrying the intact spoIIG gene restored sporulation of strain HU1002 (spoIIG41 recE4) to a frequency of 104 spores/ml and inhibited sporulation of strain 4309 (spo+ recE4) to a level of 103 spore/ml.