Willems H, Jäger C, Baljer G
Institute of Hygiene and Infectious Diseases of Animals, Justus-Liebig-Universität Giessen, D-35392 Giessen, Germany.
J Bacteriol. 1998 Aug;180(15):3816-22. doi: 10.1128/JB.180.15.3816-3822.1998.
Pulsed-field gel electrophoresis and PCR techniques have been used to construct a NotI macrorestriction map of the obligate intracellular bacterium Coxiella burnetii Nine Mile. The size of the chromosome has been determined to be 2,103 kb comprising 29 NotI restriction fragments. The average resolution is 72.5 kb, or about 3. 5% of the genome. Experimental data support the presence of a linear chromosome. Published genes were localized on the physical map by Southern hybridization. One gene, recognized as transposable element, was found to be present in at least nine sites evenly distributed over the whole chromosome. There is only one copy of a 16S rRNA gene. The putative oriC has been located on a 27.5-kb NotI fragment. Gene organization upstream the oriC is almost identical to that of Pseudomonas putida and Bacillus subtilis, whereas gene organization downstream the oriC seems to be unique among bacteria. The physical map will be helpful in investigations of the great heterogeneity in restriction fragment length polymorphism patterns of different isolates and the great variation in genome size. The genetic map will help to determine whether gene order in different isolates is conserved.
脉冲场凝胶电泳和聚合酶链反应技术已被用于构建专性细胞内细菌伯氏考克斯体Nine Mile株的NotI酶切大片段图谱。已确定该染色体大小为2103 kb,由29个NotI酶切片段组成。平均分辨率为72.5 kb,约占基因组的3.5%。实验数据支持线性染色体的存在。通过Southern杂交将已发表的基因定位在物理图谱上。发现一个被认为是转座元件的基因至少存在于整个染色体上均匀分布的9个位点。16S rRNA基因只有一个拷贝。假定的oriC位于一个27.5 kb的NotI片段上。oriC上游的基因组织与恶臭假单胞菌和枯草芽孢杆菌几乎相同,而oriC下游的基因组织在细菌中似乎是独特的。该物理图谱将有助于研究不同分离株的限制性片段长度多态性模式的巨大异质性以及基因组大小的巨大差异。遗传图谱将有助于确定不同分离株中的基因顺序是否保守。