Samuels D S, Mach K E, Garon C F
Laboratory of Vectors and Pathogens, Rocky Mountain Laboratories, National Institute of Allergy and Infectious Diseases, Hamilton, Montana 59840.
J Bacteriol. 1994 Oct;176(19):6045-9. doi: 10.1128/jb.176.19.6045-6049.1994.
No useful method to genetically manipulate Borrelia burgdorferi, the causative agent of Lyme disease, has been developed previously. We have used resistance to the coumarin antibiotic coumermycin A1, an inhibitor of DNA gyrase, as a genetic marker to monitor the transformation of B. burgdorferi by electroporation. Introduction of site-directed mutations into the gyrB gene demonstrated that transformation was successful, provided evidence that homologous recombination occurs on the chromosome, and established that mutations at Arg-133 of DNA gyrase B confer coumermycin A1 resistance in B. burgdorferi. The coumermycin A1-resistant gyrB marker and genetic transformation can now be applied toward dissecting the physiology and pathogenesis of the Lyme disease agent on a molecular genetic level.
此前尚未开发出对莱姆病病原体伯氏疏螺旋体进行基因操作的有效方法。我们利用对香豆素类抗生素香豆霉素A1（一种DNA促旋酶抑制剂）的抗性作为遗传标记，来监测通过电穿孔法对伯氏疏螺旋体的转化。将定点突变引入gyrB基因表明转化是成功的，提供了同源重组发生在染色体上的证据，并确定DNA促旋酶B的第133位精氨酸突变赋予伯氏疏螺旋体对香豆霉素A1的抗性。香豆霉素A1抗性gyrB标记和基因转化现在可用于在分子遗传学水平上剖析莱姆病病原体的生理学和发病机制。
