Ezaki B, Mori H, Ogura T, Hiraga S
Department of Molecular Genetics, Kumamoto University Medical School, Japan.
Mol Gen Genet. 1990 Sep;223(3):361-8. doi: 10.1007/BF00264441.
The seg-3 mutant Escherichia coli does not support the maintenance of mini-F plasmid at 42 degrees C. We cloned the chromosomal DNA segment of the wild-type strain W3110 that complements the Seg- phenotype of this mutant. Cleavage mapping of this segment showed that it was derived from the 76-min region of the E. coli chromosome map. Complementation tests using plasmids carrying subcloned DNA segments suggested that the seg-3 mutant carried two mutations that additively affected the maintenance of mini-F plasmid; one was in the ugpA gene and the other was presumably in the rpoH gene. We generated a disrupted ugpA null mutant and found that the mini-F plasmid was unstable in this ugpA null mutant even at 30 degrees C. This suggests that the ugpA gene product is required for the stable maintenance of mini-F plasmid.
seg-3突变型大肠杆菌在42℃时无法维持mini-F质粒。我们克隆了野生型菌株W3110的染色体DNA片段,该片段可互补此突变体的Seg-表型。对该片段的切割图谱分析表明,它源自大肠杆菌染色体图谱的76分钟区域。使用携带亚克隆DNA片段的质粒进行的互补试验表明,seg-3突变体携带两个对mini-F质粒维持有累加影响的突变;一个位于ugpA基因,另一个可能位于rpoH基因。我们构建了一个ugpA基因破坏的无效突变体,发现即使在30℃时,mini-F质粒在这个ugpA无效突变体中也不稳定。这表明ugpA基因产物是mini-F质粒稳定维持所必需的。