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Biomedical subjects

D J Kopecko

Publications and source records attributed to D J Kopecko.

61 records · Page 4Linked to original sources
PubMed1977

Transfer of plasmid-borne beta-lactamase in Neisseria gonorrhoeae.

Neisseria gonorrhoeae strain GC82 contains a plasmid specifying a beta-lactamase (beta-Lam(+)). Mixed incubation of strain GC82 with a penicillin-susceptible (beta-Lam(-)), streptomycin-resistant mutant of strain GC9 results in the expression of beta-lactamase activity and streptomycin resistance in the transcipients. The frequency of transfer of the plasmid-specified resistance to penicillin seems to be proportional to the initial input ratio of the mating mixture of donor to recipient and to correlate positively with bacterial density. Cell-to-cell transmission of the deoxyribonucleic acid (DNA) appears to be by a conjugal mechanism or, alternatively, by an as yet undescribed transducing phage. Additionally, whole-cell DNA from a beta-lactamase-producing strain could be used to transform streptomycin-resistant recipients, resulting in the expression of both beta-lactamase activity and streptomycin resistance in the transformants, and purified gonococcal plasmid DNA transformed Escherichia coli but not the gonococcus. Circular DNA extracted from donor GC82 comprised three molecular species (approximately 2.7, 4.8, and 25 megadaltons [Mdal]), whereas the recipients GC9-S (Str(r)) contained only the 2.7-Mdal cryptic DNA species. DNA from the GC9-S82 (Str(r), beta-Lam(+)) transcipient contained a 4.8-Mdal species in addition to the cryptic molecular species (2.7 Mdal). The finding that the transcipient will not retransfer beta-lactamase is consistent with the hypothesis that the 25-Mdal plasmid promotes mobilization of the smaller 4.8-Mdal R plasmid.

DNA, Bacterial↗
PubMed1977

Survey of the extrachromosomal gene pool of Streptococcus mutans.

Fifty strains of Streptococcus mutans independently isolated from human dental plaque were examined for the presence of covalently closed circular plasmid deoxyribonucleic acid (DNA). Cesium chloride-ethidium bromide centrifugation of [3H]thymidine-labeled, Sarkosyl-lysed cells revealed that 2 of the 50 strains contained plasmid DNA. The plasmid DNA from these strains was characterized by velocity and equilibrium centrifugation and by electron microscopy. The plasmids in these strains were virtually identical in size, with molecular weights of 3.6 X 10(6) and 3.7 X 10(6), Both were present to the extent of approximately 20 molecules per genome equivalent. Interlocked catenated dimeric molecules of each plasmid were readily detected by velocity sedimentation and electron microscopy. These plasmid-containing strains were compared with representative plasmid-free S. mutans strains by using such criteria as bacteriocin production, antibiotic susceptibility, and hemolysis of mammalian erythrocytes. Although no correlation of phenotype to plasmid content could be made, production of bacteriocin-like activity differed significantly between the two plasmid-containing S. mutans isolates. Thus, although the plasmids in these two isolates appeared identical by the criteria of molecular weight, presence of dimers, and copy number, they appeared to be harbored by two distinct S. mutans strains.

Anti-Bacterial Agents↗
PubMed1976

Structural evolution of bacterial plasmids: role of translocating genetic elements and DNA sequence insertions.

Recent evidence suggests that plasmids have evolved by site-specific recombinational events involving translocation and insertion of discretely defined DNA segments. The role of translocating genetic elements and repeated DNA sequences in the formation and structural evolution of bacterial plasmids, and in the control of plasmid gene expression, is the subject of this brief review. Insertion sequence (IS) regions are discrete segments of DNA that are known to cause strongly polar mutations in the genes of Escherichia coli and several bacteriophages as a consequence of their insertion into bacterial or phage genomes. Recent investigations have identified three separate kinds of IS segments on plasmids, and have indicated that such regions may have a role in 1) site-specific reversible dissociation of antibiotic resistance plasmids into their component segments, 2) recombination of certain plasmids with the bacterial chromosome, and 3) translocation of segments of plasmid DNA onto other replicons, or onto different sites of the same replicon. In addition, such DNA sequences, which may be repeated on plasmid genomes in either direct or reverse orientation, are involved in the control of plasmid gene expression. Inverted repeats other than the genetically characterized IS segments also appear to be involved in recA-independent, recombination and translocation of plasmid DNA segments. These inverted repeats contain palindromic nucleotide sequences on each strand of DNA and are detectable as hairpin-loop structures by electron microscope heteroduplex analysis. Such palindromes resemble the recognition sites for restriction endonucleases, some of which are encoded by plasmids, suggesting that similar endonucleolytic enzymes may be involved in the translocation of plasmid DNA segments.

Base Sequence↗
PubMed1975

Site specific recA--independent recombination between bacterial plasmids: involvement of palindromes at the recombinational loci.

A recA-independent recombinational event is described which results in insertion of an entire plasmid genome at a unique site of another plasmid, and coincident excision of a precisely defined DNA segment originally present at the point of the insertion. The resulting recombinant molecules subsequently can undergo site-specific translocation of their component segments or inversion of their original DNA sequence orientation. The events observed entail nonreciprocal exchange of genetic material, and involve a discrete nucleotide sequences that is duplicated in rotationally symmetrical reverse orientation on plasmid DNA (i.e., inverted repeat; palindrome).

Base Sequence↗
PubMed1972

Positive and negative control of R-factor replication in Proteus mirabilis.

Replication of the 50 and 58 moles per cent guanine plus cytosine (%GC) components of R factor 222 in Proteus mirabilis during growth in the presence and absence of chloramphenicol and after shifting exponential- and stationary-phase cells to conditions which inhibit host protein or deoxyribonucleic acid (DNA) synthesis was examined. Chloramphenicol reduced the growth rate but increased the amount of both R-factor components; the 58% GC component showed a larger proportionate increase. This was inferred to indicate reduced synthesis of an inhibitor that acts on both R-factor components and an initiator for replication of the 50% GC component. Replicative patterns observed after shifting exponential- and stationary-phase cells grown with or without chloramphenicol to minimal medium or chloramphenicol for one generation, or puromycin for 3 hr, corroborated this interpretation. After shifts of exponential cells from either medium, replication of the 50% GC components paralleled host replication, thus indicating a requirement for protein synthesis; replication of the 58% GC component increased due to reduced inhibitor synthesis. R-factor DNA remained constant after shifting stationary cells from drug-free medium, thus indicating that the cells contained effective concentrations of the regulatory inhibitor, whereas increased replication of the 58% GC component occurred after identical shifts of chloramphenicol-grown cells of the same chronological age. Similar responses were observed after shifts to 5 C or to medium containing streptomycin or tetracycline. Absence of replication of the 50% GC component after shifting to medium containing nalidixic acid or phenethanol and its hereditary persistence during growth indicated that the 50% GC replicon was attached to the membrane. Thus, in P. mirabilis the three replicons of R factor 222 are regulated as follows: The composite and transfer portion (RTF) replicons represented by the 50% GC component require protein synthesis and membrane attachment and are negatively regulated by an inhibitor; the 58% GC or resistance-determinants replicon exists cytoplasmically and is subject only to negative control.

Alcohols↗