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Genetic evolution of bacteriophage. I. Hybrids between unrelated bacteriophages P22 and Fels 2.

A new bacteriophage species, designated F22, was isolated from phage P22 stocks grown on Salmonella typhimurium Q1 lysogenic for Fels 2 at a frequency of less than 10(-11). P22 has a very short tail with a hexagonal base plate and six spikes. Phage Fels 2 is morphologically similar to E. coli T-even phages, having a long tail with a contractile sheath and carrying no genetic region related to P22. Phage F22 is morphologically and serologically indistinguishable from Fels 2, but carries the c(c(1), c(2), and c(3)) markers of P22. The color markers h(21), g, and m(3) of P22 do not appear in F22. Thus, F22 is evidently a recombinant between the unrelated bacteriophages P22 and Fels 2. The recombination between unrelated bacteriophages could play an important role in the evolution of bacteriophages.

Complement Fixation Tests↗

A mutant of Escherichia coli permitting replication of two F factors.

Control mechanisms of replication of bacterial genetic elements are poorly understood at present. We have studied one such mechanism involving replication of the F factor in Escherichia coli. The F factor can replicate either autonomously in F(+) or F' strains, or as an integral part of the chromosome in Hfr strains. We have shown that presence of either an integrated F factor or a free F factor prevents replication of a second free F factor. Two integrated F factors can replicate in the same cell. The present experiments show that when a F'lac element was transferred by mating into an Hfr strain, it had to become integrated into the chromosome in order to persist. With a recombination-deficient (recA) Hfr strain as recipient, the frequency of F'lac integration was greatly reduced. This permitted us to isolate a mutant Hfr strain in which the F'lac element was able to replicate autonomously. The mutation has most likely occurred in the integrated F factor itself. Availability of this new recA mutant Hfr strain facilitates genetic analysis of the F factor, since in recA(+) Hfr strains frequent integration of a free F factor into the chromosome obscures recognition of complementation and recombination between two free F factors.

Chromosomes, Bacterial↗

A mutant of Escherichia coli defective in DNA polymerase II activity.

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.

Chromatography↗

Gene transfer in Caulobacter crescentus: polarized inheritance of genetic markers.

Recombination frequencies were determined for 15 independently isolated auxotrophs of C. crescentus crossed pairwise in all possible combinations. The results indicate that the mutants may be grouped into at least two types: "fertile" strains, which recombine with all other mutants at frequencies ranging from less than 10-6 to 3 times 10-2, and "nonfertile" strains which recombine with fertile strains at high frequencies and with other nonfertile strains at low or negligible frequencies. Several lines of evidence indicate a polarized inheritance of markers. Two of these are (1) the preferential inheritance of unselected markers from the nonfertile parent in fertile times nonfertile crosses, and (2) the consistent ordering of markers based on the frequency at which the mutants recombine with each of the three fertile strains. Although the evidence is not conclusive at this point, the results are most consistent with conjugation at the mechanism of gene transfer in these bacteria.

Cell Division↗

Analysis of increasing antibiotic resistance of Klebsiella pneumoniae relative to changes in chemotherapy.

Three antibiotic-resistant strains of Klebsiella pneumoniae responsible for outbreaks of systemic infection in a hospital nursery were analyzed. Each organism emerged after prolonged use of a drug to which it was resistant. The first strain (RO16) produced neomycin phosphotransferase and contained three plasmids with molecular weights of 24, 25, and 30 X 10(6) daltons, respectively. Resistance to ampicillin, carbenicillin, neomycin, and kanamycin was transferred by conjugation at a frequency of 10(-6). The second strain of K. pneumoniae (RO106) produced gentamicin adenyltransferase and maintained aminoglycoside resistance only when propagated in antibiotic-containing medium. DNA analyses revealed eight species of plasmid DNA: one species with a molecular weight of 70 X 10(6) daltons apparently accounted for conjugal transfer of resistance to ampicillin, carbenicillin, chloramphenicol, and streptomycin. The third strain (RO180) was resistant primarily to colistin and lacked plasmids. Control of the outbreak due to this strain was achieved by aminoglycoside therapy.

Aminoglycosides↗