Vallari D S, Rock C O
J Bacteriol. 1985 Oct;164(1):136-42. doi: 10.1128/jb.164.1.136-142.1985.
Mutants of Escherichia coli K-12 defective in the pantothenate permease (panF) were isolated and characterized. The panF mutation resulted in the complete loss of pantothenate uptake and of the ability to use extracellular vitamin for growth. The growth phenotypes of panF panD, panF panB, and panF panC double mutants showed that the cytoplasmic membrane was impermeable to external pantothenate. Analysis of the intracellular and extracellular metabolites from strain DV1 (panF panD) labeled with beta-[3-3H]alanine demonstrated that a carrier-mediated mechanism for efficient pantothenate efflux remained in the panF mutant. Genetic mapping of this nonselectable allele was facilitated by the isolation of three independent Tn10 insertions close to panF. Two- and three-factor crosses located panF at minute 72 of the E. coli chromosome and established the gene order fabE panF aroE.
分离并鉴定了泛酸通透酶(panF)存在缺陷的大肠杆菌K-12突变体。panF突变导致泛酸摄取完全丧失,以及利用细胞外维生素进行生长的能力丧失。panF panD、panF panB和panF panC双突变体的生长表型表明,细胞质膜对细胞外泛酸是不可渗透的。对用β-[3-³H]丙氨酸标记的DV1菌株(panF panD)的细胞内和细胞外代谢物进行分析表明,panF突变体中仍存在一种载体介导的高效泛酸外排机制。通过分离三个靠近panF的独立Tn10插入片段,促进了这个非选择性等位基因的遗传定位。双因子和三因子杂交将panF定位在大肠杆菌染色体的72分钟处,并确定了基因顺序为fabE panF aroE。