Römling U, Grothues D, Bautsch W, Tümmler B
Cystic Fibrosis Research Group, Abt. Biophysikalisch Chemie. Medizinische Hochschule, Hannover, FRG.
EMBO J. 1989 Dec 20;8(13):4081-9. doi: 10.1002/j.1460-2075.1989.tb08592.x.
A complete macrorestriction map of the 5.9 Mb genome of Pseudomonas aeruginosa PAO (DSM 1707) was constructed by the combination of various one- and two-dimensional pulsed field gel electrophoresis techniques. A total of 51 restriction sites (36 SpeI sites, 15 DpnI sites) were placed on the physical map yielding an average resolution of 110 kb. Several genes encoding virulence factors and enzymes of metabolic pathways were located on the anonymous map by Southern hybridization. Distances between the gene loci were similar on the genetic and physical maps, suggesting an even distribution of genome mobility throughout the bacterial chromosome. The four rRNA operons were organized in pairs of inverted repeats. The two-dimensional macro-restriction techniques described herein are generally applicable for the genome mapping of any prokaryote and lower eukaryote which yields resolvable fragment patterns on two-dimensional pulsed field gels.
通过多种一维和二维脉冲场凝胶电泳技术的结合，构建了铜绿假单胞菌PAO（DSM 1707）5.9 Mb基因组的完整宏观限制性图谱。共有51个限制性酶切位点（36个SpeI位点，15个DpnI位点）定位在物理图谱上，平均分辨率为110 kb。通过Southern杂交，在匿名图谱上定位了几个编码毒力因子和代谢途径酶的基因。基因座之间的距离在遗传图谱和物理图谱上相似，表明基因组流动性在整个细菌染色体上分布均匀。四个rRNA操纵子以反向重复对的形式排列。本文所述的二维宏观限制性技术通常适用于任何在二维脉冲场凝胶上产生可分辨片段模式的原核生物和低等真核生物的基因组图谱绘制。
