Kuipers E J, Israel D A, Kusters J G, Blaser M J
Division of Infectious Diseases, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-2605, USA.
J Bacteriol. 1998 Jun;180(11):2901-5. doi: 10.1128/JB.180.11.2901-2905.1998.
Many strains of Helicobacter pylori are naturally competent for transformation in vitro. Since there is a high degree of genetic variation among H. pylori strains, we sought to determine whether mechanisms of DNA exchange other than transformation exist in these organisms. Studies were done with H. pylori cells that each were resistant to two different antibiotics; the procedure used involved mating of cells on plates or in broth, in the absence or presence of DNase. In each experiment, such matings produced progeny with the markers of both parents. Examination of the full resistance profile and random arbitrarily primed DNA PCR (RAPD-PCR) profiles of the progeny indicated that DNA transfer was bidirectional. DNase treatment reduced but did not eliminate transfer; only the presence of both DNase and a membrane separating the cells did so. For progeny derived from matings in the presence of DNase, antibiotic resistance and RAPD profiles indicated that transfer was unidirectional. DNase-treated cell-free supernatants also did not transform, ruling out transduction. These experiments indicate that both a DNase-sensitive mechanism (transformation) and a DNase-resistant conjugation-like mechanism involving cell-to-cell contact may contribute to DNA transfer between H. pylori cells.
许多幽门螺杆菌菌株在体外天然具有转化能力。由于幽门螺杆菌菌株之间存在高度的遗传变异,我们试图确定这些生物体中是否存在除转化之外的DNA交换机制。我们使用对两种不同抗生素具有抗性的幽门螺杆菌细胞进行了研究;所采用的程序包括在平板上或肉汤中使细胞进行交配,无论有无DNA酶。在每个实验中,这种交配都产生了具有双亲标记的后代。对后代的完整抗性谱和随机任意引物DNA PCR(RAPD-PCR)谱的检查表明,DNA转移是双向的。DNA酶处理减少了但并未消除转移;只有同时存在DNA酶和分隔细胞的膜时才会消除转移。对于在DNA酶存在下交配产生的后代,抗生素抗性和RAPD谱表明转移是单向的。经DNA酶处理的无细胞上清液也不能进行转化,排除了转导作用。这些实验表明,一种对DNA酶敏感的机制(转化)和一种涉及细胞间接触的对DNA酶有抗性的类似接合的机制可能都有助于幽门螺杆菌细胞之间的DNA转移。