Stettler R, Leisinger T
Mikrobiologisches Institut, Eidgenössische Technische Hochschule, ETH-Zentrum, Zürich, Switzerland.
J Bacteriol. 1992 Nov;174(22):7227-34. doi: 10.1128/jb.174.22.7227-7234.1992.
A physical map of the Methanobacterium thermoautotrophicum Marburg chromosome was constructed by using pulsed-field gel electrophoresis of restriction fragments generated by NotI, PmeI, and NheI. The order of the fragments was deduced from Southern blot hybridization of NotI fragment probes to various restriction digests and from partial digests. The derived map is circular, and the genome size was estimated to be 1,623 kb. Several cloned genes were hybridized to restriction fragments to locate their positions on the map. Genes coding for proteins involved in the methanogenic pathway were located on the same segment of the circular chromosome. In addition, the genomes of a variety of thermophilic Methanobacterium strains were treated with restriction enzymes and analyzed by pulsed-field gel electrophoresis. The sums of the fragment sizes varied from 1,600 to 1,728 kb among the strains, and widely different macrorestriction patterns were observed.
利用NotI、PmeI和NheI产生的限制性片段进行脉冲场凝胶电泳,构建了嗜热自养甲烷杆菌马尔堡菌株染色体的物理图谱。片段的顺序是通过NotI片段探针与各种限制性酶切产物的Southern杂交以及部分酶切产物推导出来的。推导得到的图谱是环状的,基因组大小估计为1623kb。将几个克隆基因与限制性片段杂交,以确定它们在图谱上的位置。编码参与产甲烷途径蛋白质的基因位于环状染色体的同一区段。此外,用限制性酶处理了多种嗜热甲烷杆菌菌株的基因组,并通过脉冲场凝胶电泳进行分析。菌株之间片段大小的总和在1600至1728kb之间变化,并且观察到了广泛不同的宏观限制性图谱。