Abstract
A substituted sex-factor of Escherichia coli, F'8 gal, was transferred to Proteus mirabilis by conjugation. The DNA of the episome was partially purified from Proteus DNA by preparative equilibrium centrifugation in caesium chloride, and by a bulk method using hydroxyapatite. The buoyant density of the episomal DNA is 1.707, corresponding to a (G+C) content of 47%. By optical renaturation the genetic complexity of the episomal DNA was found to be 76x10(6) daltons. RNA was synthesized in vitro by using the episomal DNA as template. By hybridizing this RNA with DNA extracted from E. coli carrying F'8 gal, it is shown that the number of copies of the episome per replicating chromosome is close to two during exponential growth. The episome makes up about 4.4% of the total DNA of the growing cells. The activities of galactokinase and galactose 1-phosphate uridylyl-transferase in cells with and without episomal and chromosomal gal genes were found to be proportional to the number of gal genes present, when the cells were induced with d-fucose, but not when they were induced with d-galactose.
MeSH Terms
Centrifugation, Density Gradient
Cesium
Chromatography
Chromosomes, Bacterial
Conjugation, Genetic
DNA Replication
DNA, Bacterial/isolation & purification
Enzyme Induction
Escherichia coli/cytology,enzymology
Extrachromosomal Inheritance
Fucose/metabolism
Galactose/metabolism
Genes
Genetics, Microbial
Hydroxyapatites
Methods
Nucleic Acid Hybridization
Nucleic Acid Renaturation
Nucleotidyltransferases/metabolism
Phosphotransferases/metabolism
Proteus
RNA/biosynthesis
Templates, Genetic
Chemicals
DNA, Bacterial
Hydroxyapatites
Cesium
Fucose
RNA
Phosphotransferases
Nucleotidyltransferases
Galactose
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Frame R
Bishop J O
References (25)
25 references, click to expand
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