Wide ranging plasmid bearing the Pseudomonas aeruginosa tryptophan synthase genes.
Explore the source record for details and available documents.
Biomedical subjects
Publications and source records attributed to R W Hedges.
Explore the source record for details and available documents.
R938 carries a transposon (TAbeta) of approximate molecular weight 9.5 Megadaltons (Mdal, 10(6) daltons). This contains genes for a beta lactamase of type TEM-1 and for streptomycin phosphatransferase (SPT). There is a ten-fold difference in the efficiency of transposition in different strains of E. coli K12.
Explore the source record for details and available documents.
Five R plasmids transferred from Asian strains of Vibrio cholerae all proved to be members of compatibility group C. A non-self-transmissible plasmid, stable in V. cholerae, was mobilized for transfer to Escherichia coli K-12 and found to be unstably inherited in that host. Plasmids of group C and P transferred to a wild V. cholerae strain were stably inherited.
Plasmids have been constructed by insertion of DNA from Rhizobium leguminosarum or Proteus mirabilis into RP4 (an R factor of group P). Such recombinant plasmids retain the wide host range of the parental plasmid, being as efficiently transmissible as the unmodified RP4 and are stably maintained in rapidly growing cultures. The recombinant plasmids, even though each contained a DNA sequence absolutely identical with that of the host strain, are no more efficient at mobilizing the transfer of chromosomal genetic information from that host strain than was unmodified RP4. We therefore conclude that an unknown factor must be essential in the process of chromosome mobilization and rate limiting for that process.
Explore the source record for details and available documents.
R factor-determined beta-lactamases have been investigated by analytical isoelectric focusing. The enzymes such as those specified by the R6K and RP4 plasmids (TEM-type enzymes) are notably homogenous in biochemical tests (Hedges et al., 1974), but two subclasses can be distinguished by isoelectric focusing. Three subclasses can be distinguished among the oxacillin-hydrolyzing enzymes, in good agreement with the classification based upon biochemical characteristics (Dale and Smith, 1974). The TEM-type beta-lactamases are promiscuously distributed among plasmids of a wide variety of compatibility groups, whereas the various oxacillin-hydrolyzing enzymes show some degree of correlation with compatibility.
R483, a plasmid of the Ialpha incompatibility group, contained a deoxyribonucleic acid (DNA) sequence encoding resistance to trimethoprim (TpR) and streptomycin (SmR) that could be transposed to other replicons, i.e., to the Escherichia coli chromosome and to related and unrelated plasmids. Each transposition resulted in the acquisition by the recipient replicon of a segment of DNA of about 9 X 10(6) daltons, both resistance genes, but never the colicin Ia or pilus genes of R483. Transposition took place at a single chromosomal site between dnaA and ilv and did not suppress the DnaA phenotype, in contrast to integration of the whole R483 plasmid. The chromosome, having received the transposition, could secondarily act as a transposition donor to another plasmid. Such a plasmid was indistinguishable from one having received a direct transposition from R483. TpR SmR transposition was very site specific and did not require a functional recA+ gene. We postulate that the TpR SmR segment of R483 is a transposon (TnC) with specific boundary sequences.
R plasmids of incompatibility group P-2 are readily transmissible between Pseudomonas strains, but not to Escherichia coli or other enterobacteria, whereas those of group P-1 have a broad host range. Pseudomonas aeruginosa donor strains carrying both a P-1 plasmid (RP1, RP4, or R751) and a P-2 plasmid (pMG1, pMG2, pMG5, or RPL11) were mated with E. coli K-12, and selection was imposed for resistance markers on the P-2 plasmids. Transconjugants were obtained at a low frequency, in which P-2 markers were expressed and were serially transmissible in E. coli together with P-1 markers. These plasmids had P-1 incompatibility properties, conferred susceptibility to phages active on P-1 carrying strains, and behaved on sucrose gradient centrifugation as unimolecular species of higher molecular weights than the P-1 parent. Recombinant plasmid formation was independent of a functional Rec gene in both donor and recipient and, with R751, had a preferred site leading to loss of trimethoprim resistance. Interaction between insertion sequences may be involved. Thus, plasmids of group P-2 can recombine with R factors of another group quite separate in compatibility properties, host range, and pilus type. Formation of such recombinants provides one pathway by which the genetic diversity of plasmids may have evolved.
Although phages P1 and PhiAmp are heteroimmune (Chesney and Scott, 1975 and Yarmolinsky, unpublished), their plasmid prophages are incompatible. Thus, the immunity and compatibility systems are two distinct regulators of phage replication. The two prophages, and plasmid P15B (Ikeda, Inzuka and Tomizawa, 1970) constitute a new compatibility group, designated Y.
Recombinant plasmids have been produced both by transduction of genetic material from FIIR factors into Proteus mirabilis strains carrying plasmids of group J and by insertion of a transposon conferring streptomycin and trimethoprim resistances into a J group R factor. The transposon-carrying derivative and one of the transductants were shown to be members of group J whereas another transductant was shown to be compatible with members of this group. This recombinant plasmid was able to eliminate but not to be eliminable by R factors of group FII. A model for the origin of this anomalous compatibility characteristic is presented based on the assumption that the recombinant plasmid carries part but not all of a complex of binding sites for the repressor of replication [Uhlin and Nordström (1975)].
A 5 Megadalton segment of DNA carrying a gene for kanamycin resistance from R447 b (a plasmid of group N of molecular weight 33 Megadaltons) has been inserted into Plac (a plasmid of the A--C complex of molecular weight 101 Megadaltons) to produce the recombinant plasmid Plac-R447 b (Coetzee, 1974). The recombinant plasmid is a typical member of the A--C complex except that entry of an N group plasmid into a Plac-R447 b+ recipient frequently leads to the loss of 5 Megadaltons of DNA (including the kanamycin resistance determinant) from the resident plasmid. In those transcipients from which kanamycin resistance is not eliminated, both plasmids are stably inherited.
RSU2, a plasmid transmissible between strains of P. aeruginosa but not to Escherichia coli can be mobilized by R751. Conjugatants receive a single plasmid composed of DNA from both R751 and RSU2 which has the compatibility properties of a member of group P (like R751). Study of this fusion plasmid suggests that the failure of RSU2 to transfer into enterobacteria is due to an inability to replicate in these bacteria. The fusion plasmid replicates using the genes of R751.
Transfer of plasmids of group S is much more efficient at low temperatures (e.g. 22 degrees C) than at 37 degrees C. This is due to failure of the donor strain to produce the transfer system during growth at the higher temperature.
In recent years, Serratia marcescens has become established in certain localities as an agent of hospital infection and cross-infection (Clayton & von Graevenitz, 1966; Wilfert, Barrett & Kass, 1968; Davis, Foltz & Blakemore, 1970; Wilkowske, Washington, Martin & Ritts, 1970). In general, strains of S. marcescens isolated from infective lesions differ from those from other sources in being non-pigmented and antibiotic resistant (Ewing, Johnson & David, 1962; Clayton & von Graevenitz, 1966). Medeiros & O'Brien (1969) and Schaefler et al. (1971) described strains of S. marcescens, isolated from hospital patients, which were able to transfer R factors to Escherichia coli. Transfer of resistance to E. coli has also been reported from strains of S. marcescens isolated in France (Grimont & Dulong de Rosnay, 1972; Scavizzi, 1972; Lemosquet-Villemon, Morel & Freymuth, 1973)9 We have collected strains of S. marcescens, most, but not all, clinical isolates, from widely separate geographical areas; each strain was tested for antibiotic resistance and for R factors transmissible to E. coli K129 The R factors were classified by compatibility in K12 (Datta, 1974). Our purpose was to find out how much of the antibiotic resistance observed in S. marcescens is characteristic of that genus and to what extent it is shared with other bacterial genera (Coetzee, Datta & Hedges, 1972; Datta & Hedges, 1972a; Hedges, 1974). The R factors described by Medeiros & O'Brien (1969) and Lemosquet-Villemon et al. (1973) were included in this study.
Eighty-nine R factors were transmitted by conjugation to Escherichia coli K12 from isolates of Proteus hauseri (P. mirabilis plus P vulgaris). More than half were non-selftranmissible. The remainder included plasmids assigned to the previously defined groups FII,A-C complex, J, N and P, as well as some not belonging to any knwon compatibility groups. R factors from strains isolated in India, Thailand and Japan carried plasmids whose inheritance was extremely unstable in E. coli K12. All belonged to a new compatibility group, V.
Explore the source record for details and available documents.
R1033 is a plasmid of compatibility group P (= P1) transferred from a wild strain of Pseudomonas aeruginosa. It confers resistance to gentamicin by gentamicin acetyl-transferase 1 and to kanamycin and neomycin by neomycin phosphotransferase 1.