Mintz C S, Shuman H A
Proc Natl Acad Sci U S A. 1987 Jul;84(13):4645-9. doi: 10.1073/pnas.84.13.4645.
Legionnaires disease is an acute respiratory disease that is often fatal for immunocompromised patients. The causative agent of this disease, Legionella pneumophila, is a Gram-negative bacterium that is present in a variety of aquatic environments. L. pneumophila is a facultative intracellular parasite; it grows within human phagocytic cells and eventually causes their destruction. In contrast to many other intracellular parasites, L. pneumophila is a Gram-negative bacterium that can be grown in standard microbiological culture medium. To determine the factors that enable this organism to enter, survive, and multiply within human mononuclear phagocytes, we chose bacteriophage Mu, a powerful genetic tool that transposes within the host cell genome, to generate insertion mutations and gene fusions in the Legionella genome. Certain derivatives of Mu are able to generate fusions between target genes and the lac operon from Escherichia coli. We have determined that although Mu is unable to attach to L. pneumophila or complete its life cycle within Legionella, it does transpose within the Legionella genome. Transposition was detected with a mini-Mu phage that carries the lac operon of E. coli.
军团菌病是一种急性呼吸道疾病,对免疫功能低下的患者往往是致命的。这种疾病的病原体嗜肺军团菌是一种革兰氏阴性细菌,存在于多种水生环境中。嗜肺军团菌是一种兼性细胞内寄生虫;它在人类吞噬细胞内生长并最终导致细胞破坏。与许多其他细胞内寄生虫不同,嗜肺军团菌是一种革兰氏阴性细菌,可以在标准微生物培养基中生长。为了确定使这种生物体能够在人类单核吞噬细胞内进入、存活和繁殖的因素,我们选择了噬菌体Mu,一种能在宿主细胞基因组内转座的强大遗传工具,用于在军团菌基因组中产生插入突变和基因融合。Mu的某些衍生物能够在靶基因与大肠杆菌的乳糖操纵子之间产生融合。我们已经确定,虽然Mu无法附着在嗜肺军团菌上或在军团菌内完成其生命周期,但它确实能在军团菌基因组内转座。用携带大肠杆菌乳糖操纵子的微型Mu噬菌体检测到了转座。