Current linkage map of Salmonella typhimurium.
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Germ-free mice were intentionally associated with drug-resistant donor strains of Escherichia coli known to carry R factors and with drug-sensitive recipient strains. In vivo transfer of R factors was observed in all experiments, involving five different donor-recipient combinations. The number of converted recipients varied, depending upon the donor-recipient combination, but in all cases it was restricted by limiting numbers of either recipient or donor strains in the digestive tract of the microbially defined mice. Converted recipients were detected in fecal material as early as 5.5 hr after mice were associated with donor and recipient bacteria. Donors, recipients, and converted recipients were detectable in the stomach, small intestine, cecum, and large intestine of the microbially defined mice and their suckling young.
Integration of the factors F(v) and F into the chromosome of a substrain of Escherichia coli K-12 has been studied. The F(v) factor is a fertility factor derived from Col V, lacking the ability to govern the production of colicin V. The derivatives of an Hfr(v) (Hfr isolated from a V colicinogenic parent) strain, PK2 (initially isolated from C600 V(+)), were shown to retain a unique bidirectional sex factor affinity locus between recA and pheA. This site shows no affinity for the E. coli K-12 F factor as shown by inability to isolate Hfr strains with origins in this region from a parental strain containing a cytoplasmic F factor. However this area exhibits two regions of homology to the V colicinogenic factor. One gives rise to Hfr(v) strains identical to the original Hfr(v) strain, PK2, with an origin and polarity of transfer designated pheA-CC injecting markers in the order pheA-his-trp-pro. The second gives rise to strains apparently originating at the same site but with reverse polarity designated recA-C, transferring markers in the order recA-thyA-str-xyl. For strains possessing the F(v) factor only the second homology is apparent. A model for the evolution of these strains is presented.
The chromosomal location of Group I pyridoxine mutations in Escherichia coli is shown to be adjacent to dsdA,aroC, and purF (old purC) in E. coli B x K-12 hybrids. All mutants previously classified into Group I by nutrition tests and transduction frequency tests are shown to be linked to dsdA.
Mutants of Escherichia coli K-12 unable to excise pyrimidine dimers from their deoxyribonucleic acid (DNA) because of a uvr mutation show a higher survival when plated on a minimal salts medium after exposure to ultraviolet radiation than when plated on a complex medium such as nutrient agar containing yeast extract. This response has been called minimal medium recovery (MMR). Recovery of uvr mutants can take place in liquid as well as on solid medium, but not in buffer or under conditions of amino acid starvation that do not permit cell growth and normal DNA replication. MMR can thus be distinguished from the recovery of recombination-deficient (rec(-)uvr(+)) derivatives of K-12 which can occur under conditions where growth is not possible. Because MMR is characteristic of excision-defective mutants, it evidently reflects a type of repair independent of excision. We have obtained genetic evidence that MMR is determined by the rec genes, which also control recombination in K-12. Cells carrying a uvr mutation together with recA13, recA56, recB21, or recC22 failed to show MMR and were more sensitive to ultraviolet radiation than either their rec(+)uvr(-) or rec(-)uvr(+) parents. The rec(+)uvr(-) derivatives obtained from recA uvr(-) strains by transduction or by reversion regained the capacity for MMR. Our results indicate that inactivation of any one of the three genes, recA, recB, or recC, prevents cells from showing MMR.
The similarity of sex pili mediated by F factors and R(fi(+)) factors and the ability of R(fi(+)) factors to control by repression the functioning of pilus genes encoded by the F factor suggested that F factors and R(fi(+)) factors are closely related. Further comparisons of the episomal properties of F factors and R(fi(+)) factors, however, indicated many differences. F factors contain information for a restriction system for phages phiII and T7. Cells containing R factors are sensitive to these phages. Furthermore, R(fi(+)) factors do not repress the F factor phiII restriction system in cells containing both an R(fi(+)) factor and an F factor. R factors and F factors are heteroimmune episomes. In addition, an R(fi(+)) factor in cells containing both an R factor and an F factor does not fully repress the expression of F-factor immunity to an incoming second F factor. R-factor and F-factor replication systems are not identical. Wild-type F-factor replication genes will complement the mutant F(ts114)lac(+) replication genes in cells containing two F factors. The F(ts114)lac(+) episome is retained when these cells are grown at 42 C; however, cells containing an R(fi(+)) factor and F(ts114)lac(+) lose the F(ts114)lac(+) when grown at 42 C, at the same rate as cells containing only the F(ts114)lac(+). The replication system of the R(fi(+)) factor will not complement the mutant F(ts114)lac(+) replication system.
A class of mutants of Ustilago maydis selected on a fungitoxic oxathiin lack of antimycin A-tolerant respiratory system which is present in wild-type cells. This system provides, directly or indirectly, for considerable resistance to antimycin A because growth of mutant cells lacking the system is much more sensitive to the antibiotic than that of the wild type. Antimycin A-sensitive O(2) uptake and growth is found in half of the progeny from crosses of mutant to wild type. All antimycin A-sensitive segregants are somewhat more resistant to oxathiins than the antimycin A-resistant segregants. The respiration of the mutant is strongly inhibited by cyanide and azide at concentrations which stimulate respiration of the wild type. Respiration of both mutant and wild type is about equally inhibited by rotenone. It appears that the mutation alters some component of the respiratory system located between the rotenone inhibition site and the antimycin A inhibition site that permits shift of electron transport to an alternate terminal oxidase when the normal electron transport pathway is blocked.
A host cell reactivation-negative mutation, uvrD3, in Escherichia coli K-12 is located between ilv and metE.
Recombinants obtained from matings of Nocardia erythropolis x N. canicruria were tested for their genetic stability by comparing phenotypes from direct selection with the same population after unselected growth. Contraselective loci were employed in various combinations in order that all of the mapped characters might be subjected to unselected analysis. Some recombinant class types appeared as stable haploids, whereas others behaved as heterozygous diploids, segregating out new phenotypes. All regions of the parental genomes were found to be involved in segregation, implying that the entire mapped region can become merozygotic under standard mating conditions. On the basis of segregating phenotypes, the genetic potentials of these compatible nocardiae were ascertained as follows: the formation of a diploid with subsequent segregation of parental or haploid recombinant genomes or both; persistence of the diploid through many generations; continuing reassortment of genetic information by multiple matings between parental or recombinant organisms; and, very probably, second-round recombinations within the diploid. A considerable difference in the nuclear division time between the parental organisms was postulated to have significant effects on the nature of the unselected segregants.
purB and ctr mutants of Escherichia coli are resistant to low levels of nalidixic acid.
Immobilized deoxyribonucleic acid (DNA) preparations from eight species of mycobacteria were reacted with labeled reference DNA from Mycobacterium tuberculosis and M. kansasii. All mycobacteria showed some degree of homology. Immobilized DNA from Pseudomonas multivorans, which has a guanine plus cytosine content within the range established for the mycobacteria, showed no significant binding with either reference system. Relative per cent binding and thermal stability of bound DNA were used as parameters for determining degrees of relatedness among members of the genus. When these relationships were compared with those established by numberical taxonomic methods, a high correlation was found.
Mutants of Escherichia coli K-12 defective in enzyme I or Hpr, the two common components of the phosphoenolpyruvate-dependent phosphotransferase system, were isolated by a simple, direct method. The ptsI locus, the structural gene for enzyme I, and the ptsH locus, the site of mutations leading to loss of Hpr activity, are adjacent genes and could be part of a single operon. These two genes lie between the purC and supN markers in the order: strA... guaB-purC-ptsI-ptsH-supN-dsdA... his.
Genetic studies were undertaken on 14 pleiotropic negative sporulation mutants. These mutants (spoA) which are blocked early in the sporulation process were found to map near the terminus of the Bacillus subtilis chromosome in a region enriched in genes involved in spore formation. Two- and three-factor crosses by transduction and transformation led to the conclusion that the pleiotropic spoA mutations formed a linked cluster. The genetic distance across the cluster calculated from transformation data was compatible with the mutant sites defining a single gene. Suppressor studies revealed that either a nonsense or missense mutation in the spoA locus generated a pleiotropic negative phenotype. It was concluded that the locus codes for a protein, and the absence of this protein is responsible for the pleiotropic phenotype.
The genetic homogeneity among eight cariogenic strains of Streptococcus mutans was assessed by deoxyribonucleic acid (DNA)-DNA reassociation experiments. DNA species were extracted from strains GS5, Ingbritt, 10449, FAl, BHT, E49, SLl, and KlR. Labeled DNA ((14)C-DNA) was extracted from strains 10449, FAl, and SLl. Denatured (14)C-DNA fragments were allowed to reassociate, i.e., form hybrid duplexes, with denatured DNA immobilized on membrane filters incubated in 0.45 m NaCl-0.045 m sodium citrate at 67 or 75 C. At 67 C, 10449 (14)C-DNA reassociated extensively only with GS5 and Ingbritt DNA. FAl (14)C-DNA hybridized extensively only with BHT DNA, and SLl (14)C-DNA reassociated with KlR and E49 DNA. DNA which hybridized extensively at 67 C also reassociated to a high degree at 75 C. Thermal elution of (14)C-FAl-BHT duplexes showed that the hybrid duplexes were thermostable. The results indicate that S. mutans is a genetically heterogeneous species. The strains studied can be divided into three (possibly four) genetic groups, and these groups closely parallel antigenic groups.
The deoxyribonucleic acid of F-like plasmids appear to share a high degree of nucleotide similarity with each other but are not highly related to I-like plasmids.
Pseudomonas aeruginosa displays a native resistance to a variety of inhibitory compounds, including many analogues of amino acids, purines, and pyrimidines. Therefore, it has been difficult to isolate analogue-resistant regulatory mutants which have been so valuable in other microbial species for the study of enzyme control mechanisms and for the study of amino acid transport and its regulation. However, we have found that increased sensitivity to growth inhibition by analogues can be demonstrated by manipulation of the nutritional environment. When P. aeruginosa is grown with fructose as the nutritional source of carbon and energy, the cells become sensitive to growth inhibition by beta-2-thienylalanine and p-amino-phenylalanine, analogues of phenylalanine and tyrosine, respectively. Thus, mutants were isolated which are resistant to growth inhibition by beta-2-thienylalanine and p-amino-phenylalanine when fructose is the carbon source, and many of the beta-2-thienylalanine-resistant mutants overproduce phenylalanine. Several lines of evidence suggest that the increased sensitivity to growth inhibition by analogues of phenylalanine and tyrosine reflects a decreased rate of synthesis of aromatic amino acids or their precursors when fructose is the carbon source. This general approach promises to be valuable in the study of regulatory phenomena in microorganisms which, like P. aeruginosa, are naturally resistant to many metabolite analogues.