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Biomedical subjects

J N Coetzee

Publications and source records attributed to J N Coetzee.

At least 55 records · Page 3Linked to original sources
PubMed1975

Chromosome transfer in Proteus mirabilis mediated by hybrid plasmid.

A previously-described fused plasmid, P-lacRIdrd19, was found to mediate chromosomal transfer between cells of Proteus mirabilis strain PM5006; PM5006-(P-lacRIdrd19) was usually the donor and various auxotrophs of PM5006 resistant to nalidixic acid and/or streptomycin were recipients. The donor was usually counterselected with nalidixic acid and/or high concentrations of streptomycin. Recombination experiments with single markers indicated a 40-fold variation in recombination frequencies for different markers. Mapping double-auxotrophic markers by their gradient of transmission confirmed this variation and placed each of two independent isolates of eight markers in a linkage group his-ser-ura-pyrB-trp-cys-ade-ilv. Some donor markers did not register. Despite low recombination frequencies, interrupted mating experiments showed a polarity of early marker transfer. The segregation of unselected markers confirmed the order of some markers and showed that genetic material passed from the presumptive donor to the recipient. Recipients with two auxotrophic markers which could not be cotransduced by phage 5006M were converted to prototrophy by conjugation. The plasmid transferred to recipients at high frequency and all recombinants carried it. Recombinants could act as donors in further matings. Recombinants were fully susceptible to phage 5006M, unlike transductants of PM5006 by this phage. Direct involvement of the plasmid was indicated by drastically diminished recombination frequencies in crosses with recipients carrying P-lac as resident. P-lac had previously been shown to reduce the frequency of transfer of the hybrid plasmid to cells harbouring it. The histidine region was the first to register in recipients and recombined at the highest frequency of 5 times 10-6/donor cell. Some temporary association of plasmid and perhaps only the histidine region of the chromosome is favoured as the mechanism of chromosomal transfer. This could explain why not all donor markers could be mapped. Transduction and transformation were excluded as the cause of results.

Chromosome Mapping↗
PubMed1975

Specialized transduction of kanamycin resistance in a Providence strain.

Properties of a transducing system with a phage able to transduce a kanamycin-resistance marker of the T compatibility group plasmid R394 at a frequency of 2 times 10(-2)/plaque-forming unit adsorbed are described. The phage was detected in Providence strain P29 transduced to kanamycin resistance by Providence phage PL25 grown on this strain harbouring the R factor. Four P29 transductants, specially selected at the lowest multiplicities of infection of the high frequency transducing (HFT) phage, were defective lysogens. They plated PL25 with an efficiency of I and only one liberated low-titre phage spontaneously or on u.v. induction. The defect in maturation function could be corrected by introduction of a wild PL25 prophage. The transducing phage was serologically frequency was increased by the simultaneous presence of homologous non-transducing phage. Transductants did not transfer the kanamycin-resistance marker by conjugation, and produced kanamycin-sensitive segregants at a moderate rate. These segregants could be transduced to kanamycin resistance by the HFT phage. Irradiation of HFT lysates by u.v. produced an exponential fall in transduction frequency. It was concluded that the defective phage transduced by lysogenization. Kanamycin-resistant transductants could themselves be transduced by streptomycin resistance by PL25 reared on a streptomycin-resistant mutant. Lysogenic transductants produced by the HFT phage did not always liberate HFT phage on u.v. induction. Possible explantations are considered.

Bacteriophages↗
PubMed1975

Transduction of a Proteus vulgaris strain by a Proteus mirabilis bacteriophage.

Only Proteus vulgaris strain PV127 out of many P. vulgaris, P. morganii and Providence strains was transduced to kanamycin resistance by high-frequency transducing variants, 5006MHFTk and 5006MHFTak, of phage 5006M, a general transducing phage for P. mirabilis strain PM5006. The phages adsorbed poorly to strain PV127 and did not form plaques. The transduction frequency of PV127 by these phages was 5 x 10(-8)/p.f.u. adsorbed. Phage 5006M increased the transduction frequencies. Abortive transductants were not detected. Transductants segregated kanamycin-sensitive clones at high frequency and this, together with data from the inactivation of transducing activity of lysates by ultraviolet irradiation, indicated that transduction was by lysogenization. The general transducing property of the phages was not expressed in transductions to auxotrophs of PV127. Transductants (type I) resulting from low multiplicities of phage input adsorbed phage to the same extent as PV127. This suggested a defect in the transducing particles (or host) because single phage 5006M infection converted strain PM5006 to non-adsorption of homologous phage. Type I transductants did not liberate phage, suggesting a defective phage maturation function. Transductants (type II) which arose from higher multiplicities of phage input did not adsorb phage, indicating possible heterogeneity among transducing particles. Phage derived from type II transductants adsorbed poorly to PV127 and transduced it to kanamycin resistance at frequencies similar to those of phages 5006MHFTk and 5006MHFTak, ruling out host-controlled modification as a cause of the low transduction frequencies. This phage transduced PM5006 to antibiotic resistance at high frequencies but generalized transduction was again not detected. It was suggested that general transduction could be performed by particles which, due to a different composition and/or mode of chromosomal integration, made material they carried susceptible to host-cell modification.

Bacteriophages↗