Kadner R J, Shattuck-Eidens D M
J Bacteriol. 1983 Sep;155(3):1052-61. doi: 10.1128/jb.155.3.1052-1061.1983.
The Escherichia coli transport system responsible for the accumulation of a number of sugar phosphates is encoded by the uhp region and is induced by external, but not intracellular, glucose 6-phosphate. To delineate the genetic organization of the uhp region, a total of 225 independent point, deletion, and transposon Tn10 insertion mutations were collected. Mutations conferring the Uhp-phenotype were obtained on the basis of their resistance to fosfomycin and their inability to use sugar phosphates as carbon source. Deletions of uhp sequences were obtained as a consequence of imprecise excision of Tn10 insertions located on either side of uhp. Conjugal crosses between these deletions and the point of insertion mutations allowed determination of the relative order of the uhp alleles and of the deletion endpoints. Specialized lambda transducing phages carrying a uhpT-lac operon fusion and various amounts of adjacent uhp material were isolated and used as genetic donors. Results from these crosses corroborated those obtained in the conjugal crosses. The locations of the mutant alleles were compared with the regulatory properties of Uhp+ revertants of these alleles. This comparison suggested the existence of at least three genes in which mutation yields the Uhp-phenotype. Mapping experiments were consistent with the gene order pyrE-gltS-uhpTRA-ilvB, where uhpT encodes the transport system and uhpR and uhpA are regulatory genes whose products are necessary for proper uhp regulation.
负责多种磷酸糖积累的大肠杆菌转运系统由uhp区域编码,并由细胞外而非细胞内的6-磷酸葡萄糖诱导。为了描绘uhp区域的基因组织,共收集了225个独立的点突变、缺失突变和转座子Tn10插入突变。基于对磷霉素的抗性以及无法利用磷酸糖作为碳源,获得了具有Uhp表型的突变。由于位于uhp两侧的Tn10插入的不准确切除,导致了uhp序列的缺失。这些缺失与插入点突变之间的接合杂交允许确定uhp等位基因的相对顺序和缺失端点。分离出携带uhpT-lac操纵子融合和各种数量相邻uhp物质的专门λ转导噬菌体,并用作遗传供体。这些杂交的结果证实了在接合杂交中获得的结果。将突变等位基因的位置与这些等位基因的Uhp +回复体的调节特性进行了比较。这种比较表明存在至少三个基因,其中突变产生Uhp表型。定位实验与基因顺序pyrE-gltS-uhpTRA-ilvB一致,其中uhpT编码转运系统,uhpR和uhpA是调节基因,其产物对于适当的uhp调节是必需的。