Ayaki H, Kobayashi Y
J Bacteriol. 1984 May;158(2):507-12. doi: 10.1128/jb.158.2.507-512.1984.
Two specialized transducing phages carrying a sporulation gene, spoIIG , of Bacillus subtilis were constructed from B. subtilis temperate phages p11 and phi 105 by the "prophage transformation" method. Restriction enzyme analysis and transformation experiments showed that the spoIIG gene was present on a 6.2 X 10(6)-dalton (6.2-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 10(4) spores per ml and inhibited sporulation of strain 4309 ( spo + recE4 ) to a level of 10(3) spores per ml.
通过“原噬菌体转化”方法，从枯草芽孢杆菌温和噬菌体p11和phi 105构建了两种携带枯草芽孢杆菌芽孢形成基因spoIIG的特异性转导噬菌体。限制性内切酶分析和转化实验表明，spoIIG基因存在于两种转导噬菌体基因组中一个6.2×10⁶道尔顿（6.2 Md）的EcoRI片段上。进一步分析表明，spoIIG⁺转化活性存在于6.2 Md EcoRI片段内一个2.25 Md的EcoRI - BamHI片段上。将该2.25 Md片段亚克隆到pUB110的EcoRI和BamHI位点之间的区域，并构建了在2.25 Md片段内缺失PstI或HindIII片段的缺失质粒。携带完整spoIIG基因的重组质粒将HU1002菌株（spoIIG41 recE4）的芽孢形成恢复到每毫升10⁴个芽孢的频率，并将4309菌株（spo⁺ recE4）的芽孢形成抑制到每毫升10³个芽孢的水平。
