Butler P D, Moxon E R
Department of Paediatrics, University of Oxford, UK.
J Gen Microbiol. 1990 Dec;136(12):2333-42. doi: 10.1099/00221287-136-12-2333.
Contour-clamped homogeneous electric field pulsed-field gel electrophoresis (PFGE) was used in combination with Southern hybridization to construct a genomic restriction map for the pathogen Haemophilus influenzae serotype b, strain Eagan. Sites for four restriction endonucleases, EagI, NaeI, RsrII and SmaI, were located on the 2100 kbp circular chromosome. Twelve potential virulence loci have been placed on the map together with certain loci essential for growth of the bacteria (e.g. ribosomal RNA operons). PFGE also provided a valuable tool for characterizing ten capsulated, type b isolates (other than Eagan) known to be genetically heterogeneous and two laboratory-derived variants (transformants) derived through complex recombinational events involving random uptake of high-molecular-mass donor genomic DNA.
轮廓夹钳均匀电场脉冲场凝胶电泳(PFGE)与Southern杂交相结合,用于构建b型流感嗜血杆菌伊根菌株病原体的基因组限制性图谱。四种限制性内切酶EagI、NaeI、RsrII和SmaI的位点位于2100 kbp的环状染色体上。十二个潜在的毒力基因座以及细菌生长所必需的某些基因座(如核糖体RNA操纵子)已定位在该图谱上。PFGE还为鉴定十种已知基因异质性的b型包膜分离株(伊根菌株除外)和两种通过涉及随机摄取高分子量供体基因组DNA的复杂重组事件衍生的实验室变体(转化体)提供了一个有价值的工具。