Pullinger Gillian D, Bevir Thomas, Lax Alistair J
Department of Microbiology, Dental Institute, King's College London, London, UK.
Mol Microbiol. 2004 Jan;51(1):255-69. doi: 10.1046/j.1365-2958.2003.03829.x.
Toxigenic strains of Pasteurella multocida produce a 146 kDa toxin (PMT) that acts as a potent mitogen. Sequence analysis of the structural gene for PMT, toxA, previously suggested it was horizontally acquired, because it had a low G + C content relative to the P. multocida genome. To address this, the sequence of DNA flanking toxA was determined. The sequence analysis showed the presence of homologues to bacteriophage tail protein genes and a bacteriophage antirepressor, suggesting that the toxin gene resides within a prophage. In addition to phage genes, the toxA flanking DNA contained a homologue of a restriction/modification system that was shown to be functional. The presence of a bacteriophage was demonstrated in spent medium from toxigenic P. multocida isolates. Its production was increased by mitomycin C addition, a treatment that is known to induce the lytic cycle of many temperate bacteriophages. The genomes of bacteriophages from three different toxigenic P. multocida strains had similar but not identical restriction profiles, and were approximately 45-50 kb in length. The prophages from two of these had integrated at the same site in the chromosome, in a tRNA gene. Southern blot analysis confirmed that these bacteriophages contained the toxA gene.
多杀性巴氏杆菌的产毒菌株可产生一种146 kDa的毒素(PMT),该毒素是一种强效有丝分裂原。先前对PMT结构基因toxA的序列分析表明,它是水平获得的,因为相对于多杀性巴氏杆菌基因组,其G + C含量较低。为了解决这个问题,测定了toxA侧翼的DNA序列。序列分析显示存在噬菌体尾蛋白基因和噬菌体抗阻遏物的同源物,这表明毒素基因位于原噬菌体中。除了噬菌体基因外,toxA侧翼DNA还包含一个功能正常的限制/修饰系统的同源物。在产毒多杀性巴氏杆菌分离株的用过的培养基中证实了噬菌体的存在。添加丝裂霉素C可增加其产生,丝裂霉素C是一种已知可诱导许多温和噬菌体裂解周期的处理方法。来自三种不同产毒多杀性巴氏杆菌菌株的噬菌体基因组具有相似但不完全相同的限制性图谱,长度约为45 - 50 kb。其中两种的原噬菌体已整合到染色体上的同一个位点,即一个tRNA基因中。Southern印迹分析证实这些噬菌体含有toxA基因。