Evidence for a Composite State of an F'his, Gnd Element and a Cryptic Plasmid in a Derivative of Salmonella Typhimurium LT2

A method designed to select mutants constitutive for expression of the histidine operon has been applied to a Salmonella typhimurium LT2 strain containing an F'his,gnd element and a cryptic plasmid. One of the mutants isolated, strain AA0019, has not only increased levels of histidinol phosphate phosphatase (hisB), but also increased levels of gluconate-6-phosphate dehydrogenase (gnd). Ultracentrifugation studies of extrachromosomal deoxyribonucleic acid (DNA) isolated from strain AA0019 revealed the presence of a single species of covalently closed circular (CCC) DNA that sedimented more rapidly through neutral and alkaline sucrose gradients than any of its possible plasmid precursors. From neutral sucrose gradients, sedimentation coefficients of 130, 100, and 86S were derived, corresponding to the CCC DNA of the large plasmid in strain AA0019, the F'his,gnd element and the cryptic LT2 plasmid, respectively. An Escherichia coli plasmid-free strain that upon mating had received the large 130S plasmid also contained 86S and 100S CCC DNA components. A histidine-requiring derivative of strain AA0019 obtained after acridine orange treatment retained the cryptic plasmid DNA. Apparently, the large plasmid in strain AA0019 consists of the F'his,gnd element and the cryptic LT2 plasmid of the parental strain.