[Sodium and potassium in erythrocytes (author's transl)].
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Biomedical subjects
Publications and source records attributed to Y Hirota.
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A mutant of E. coli defective in DNA polymerase II activity was isolated. Extracts had about 0.1% of the normal activity of the DNA polymerase. The effect of the mutation on the ability of cells to replicate DNA of various sources was analyzed. Mutant bacteria grow normally, at 41 degrees as well as at 30 degrees . All bacteriophages, F factors, and R factors examined so far grow normally in the mutant. Sensitivity of the mutant to ultraviolet radiation and alkylating reagents in growth media was the same as that of the wild type. The mutation, polB1, is recessive with respect to wild type. The polB1 mutation can coexist with recombination-defective mutations. Genetic mapping studies show the mutation to be located at about 2 min on the E. coli map.
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The temperature-sensitive mutant CRT46 of Escherichia coli K12 is unable to initiate new rounds of DNA replication at 42 degrees C. Mutants of bacteriophage P2 have been isolated, which, in the prophage state, allow mutant CRT46 to grow at 42 degrees C. The lysogenic bacteria that grow at 42 degrees C are apparently replicating under the control of prophage P2, which substitutes for the bacterial initiation system. The ability of prophage P2 cause this suppression phenomenon depends on the position of the prophage on the bacterial chromosome. Those lysogenic strains that are able to grow at 42 degrees C all carry the prophage close to metE. The P2 mutants that allow CRT46 to grow at 42 degrees C have insertions in the early region of the P2 genome. The suppression requires the cis-acting protein formed by gene A of P2.
A series of double mutants carrying one of the thermosensitive mutations for DNA synthesis (dnaA, B, C, D, E, F, and G) and the polA1 mutation of DeLucia and Cairns, were constructed. Enzyme activities of DNA Polymerases II and III were measured in each mutant. DNA Polymerase II activity was normal in all strains tested. DNA Polymerase III activity is thermosensitive specifically in those strains having thermoscnsitive mutations at the dnaE locus. From these results we conclude that DNA Polymerases II and III are independent enzymes and that DNA Polymerase III is an enzyme required for DNA replication in Escherichia coli.
Transduction by Plkc of drug-resistance markers of the factor R213 was shown to occur at an exceptionally low frequency (at less than 10(-8) of the input phage), and they could not be transduced by P22. When the recipient cells carried a homologous R factor derived from R213, markers were transduced by Plkc at a normal frequency (at about 10(-5) to 10(-6) of the input phage). Derivative R factors, transducible by Plkc at a normal frequency but being transferred by conjugation at a frequency lower than that of the original R213, were obtained. This type of transductant often segregated R(-) cells. In addition, several transductants contained R factors which were transferred normally by conjugation but were transduced by Plkc at as low a frequency as the original R213. This type of transductant was an effective recipient for transduction by Plkc of R213 when apparently "cured" by acridine treatment. No such effective "cured" recipients were obtained from the transductants with derivatives of R213 transducible at a normal frequency. Two possible interpretations are presented: (i) R213 produces a bacteriocin-like substance upon transduction, or (ii) the genome size of R213 is too large for all of its determinants to be transduced.
A technique allowing chain formation by Escherichia coli for the analysis of the segregation of certain cellular components by autoradiographic methods has been developed. Deoxyribonucleic acid labeled with (3)H-thymidine does not appear to segregate preferentially with one or another of the bacteria. Membrane lipids labeled with either (3)H-oleate or (3)H-glycerol are evenly distributed among all progeny cells. A similar pattern of distribution of cell wall material is observed when (3)H-diaminopimelate is used as the marker.
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