von Both U, Otten S, Darbouche A, Domann E, Chakraborty T
Institut für Medizinische Mikrobiologie, Justus-Liebig-Universität, Giessen, Germany.
FEMS Microbiol Lett. 1999 Jun 15;175(2):281-9. doi: 10.1111/j.1574-6968.1999.tb13632.x.
Listeria monocytogenes is a facultative intracellular pathogen responsible for both invasive and non-invasive food-borne illness in animals and humans. In this study, macrorestriction analysis following pulsed-field gel electrophoresis was used to show that Listeria monocytogenes serovar 1/2a strain EGD has a single chromosome containing eight NotI fragments of 1100, 850, 365, 320, 275, 40, 30 and 20 kb in size and 11 AscI fragments of 860, 470, 410, 360, 320, 250, 110, 80, 50, 30 and 20 kb. The total genome therefore comprises 3000 +/- 50 kb. The creation of a physical and genetic map of the Listeria genome was achieved by generating NotI linking clones and their use in subsequent hybridisation analysis. Using isogenic mutants harbouring additional artificial NotI restriction sites, we were able to precisely map the positions of all currently known virulence genes on the chromosome.
单核细胞增生李斯特菌是一种兼性胞内病原体，可导致动物和人类发生侵袭性和非侵袭性食源性疾病。在本研究中，采用脉冲场凝胶电泳后的宏观限制性分析表明，单核细胞增生李斯特菌血清型1/2a菌株EGD有一条单一染色体，包含8个大小分别为1100、850、365、320、275、40、30和20 kb的NotI片段以及11个大小分别为860、470、410、360、320、250、110、80、50、30和20 kb的AscI片段。因此，整个基因组约为3000 +/- 50 kb。通过构建NotI连接克隆并将其用于后续杂交分析，完成了李斯特菌基因组的物理图谱和遗传图谱的构建。利用携带额外人工NotI限制性位点的同基因突变体，我们能够精确绘制出所有目前已知毒力基因在染色体上的位置。
