Plasmid identification using specific endonucleases

Thompson, Russell; Hughes, Stephen; Broda, Paul

doi:10.1007/bf00264835

Cited by 95 publications

(40 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These enzymes have been shown to be particularly useful for the analysis and identification of plasmid DNA (37). Thus, plasmids which are known to be very closely related have been readily differentiated by using restriction endonucleases and agarose electrophoresis (37). However, this technique has seen very limited use in the examination of R plasmids of clinical significance (9, 11).…”

Section: Discussionmentioning

confidence: 99%

“…Plasmid DNA from E. coli transconjugants of each strain was purified, and a molecular weight of 58 x 106 was determined for each plasmid. An analysis of the EcoRI restriction endonuclease fragments of plasmid pBP4, which had originated in a strain of K. pneumoniae type 30 isolated at the peak of the epidemic in September 1975 (Table 1 (37) cannot rule out the occurrence of minor changes confined to individual DNA fragments; but to do this unequivocally would require the sequencing of each plasmid.…”

Section: Discussionmentioning

confidence: 99%

“…However, other work-VOL. 15 ers have found the sum of restriction endonuclease fragment sizes to be 5 to 10% less than the known molecular weight of the intact plasmid (37).…”

Section: Antimicrob Agents Chemothermentioning

confidence: 99%

See 2 more Smart Citations

Physical Characterization of Ten R Plasmids Obtained from an Outbreak of Nosocomial Klebsiella pneumoniae Infections

Sadowski

Peterson

Gerding

et al. 1979

Antimicrob Agents Chemother

View full text Add to dashboard Cite

Gentamicin resistance in Klebsiella pneumoniae involved in an outbreak at the Minneapolis Veterans Administration Hospital was due to a transmissible R plasmid. In addition to gentamicin, this plasmid conferred resistance to tobramycin, kanamycin, ampicillin, carbenicillin, cephalothin, chloramphenicol, and sulfathiazole. R plasmids which transferred this complex antibiogram were identified in several clinical isolates, including four different serotypes of K. pneumoniae, Escherichia coli, Enterobacter cloacae , and Proteus morganii . The covalently closed circular form of all R plasmids isolated had a sedimentation coefficient of 76S to 77S, corresponding to a molecular weight of 58 × 10 6 . The possibility that a single R plasmid was responsible for the dissemination of multiple drug resistance among all of these different clinical strains was examined by characterizing the plasmids by using Eco RI restriction endonuclease. The same 15 fragments were obtained from each of the 10 plasmids analyzed. Their molecular weights ranged from 4 × 10 5 to 11 × 10 6 . Thus, we conclude that each of the 10 plasmids present in the various clinical strains isolated from the hospital over a 7-month period originated from a common source and that R plasmid transfer was important in their spread.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Physical Characterization of Ten R Plasmids Obtained from an Outbreak of Nosocomial Klebsiella pneumoniae Infections

Sadowski

Peterson

Gerding

et al. 1979

Antimicrob Agents Chemother

View full text Add to dashboard Cite

show abstract

“…However, the exchange of DNA within plasmids, both by acquisition and loss of transposons, and by gene mutation and deletions (24) results in plasmids of the same incompatibility group having diverse total DNA. Various studies have demonstrated DNA variability within incompatibility groups by restriction endonuclease digests and DNA hybridization (25)(26)(27)(28). While plasmid incompatibility provides evidence for a common origin, a high degree of DNA homology demonstrated by identical or similar digest patterns is a useful indicator of current identity.…”

Section: Discussionmentioning

confidence: 99%

Restriction endonuclease characterization of resistant plasmids in Enterobacteriaceae isolated from children in the Sudan

Shears

Suliman

1989

Epidemiol. Infect.

View full text Add to dashboard Cite

SUMMARYThe investigation of plasmid similarity is an important component in the surveillance of antimicrobial resistance and in the detection of epidemic plasmids. The use of restriction endonucleases in the classification of transferable, multiplyresistant plasmids from faecal Enterobacteriaceae isolated at the Children's Emergency Hospital, Khartoum was investigated. Twenty-four transconjugant plasmids, coding for 11 different resistance patterns, each of molecular weight 62 MDa, were studied using four restriction enzymes; Pst I, EcoR I, Hind III and Ava II. Fifteen different digest profiles were obtained. Restriction profiles discriminated between plasmids with differing resistance patterns and demonstrated homology of plasmids with common resistance patterns. Restriction endonuclease digest patterns provide a potentially rapid and reproducible method of plasmid classification, that could contribute towards surveillance systems in tropical countries with a high prevalence of antimicrobial resistance.

show abstract

“…S. cerevisiae DNA was purified by chloroform-isoamyl alcohol extraction of lysed spheroplasts (8). Plasmid DNA from E. coli was purified by cesium chloride-ethidium bromide centrifugation (26). Restriction enzymes were purchased from Bethesda Research Laboratories and used according to the manufacturer's instructions.…”

Section: Methodsmentioning

confidence: 99%

Recombinational instability of a chimeric plasmid in Saccharomyces cerevisiae.

Whiteway¹,

Ahmed²

1984

Mol. Cell. Biol.

View full text Add to dashboard Cite

Wild-type strains of Saccharomyces cerev'isiae exhibit mitotic recombination between the chimeric plasmid TLC-1 and the endogenous 2,u circle that involves sequence homologies between the two plasmids that are not acted on by the 2,u circle site-specific recombination system. This generalized recombination can be detected because it separates the LEU2 and CAN! markers of TLC-1 from each other through the formation of a plasmid containing only the S. cerev'isiae LEU2 region and the 2,u circle. This derivative plasmid is maintained more stably during vegetative growth than TLC-1, and strains which carry it frequently lose the endogenous 2,u circle. Therefore, TLC-1 can provide a convenient selection for [cir°] cells. Formation of this new plasmid is greatly reduced, but not eliminated, in strains containing the rad52-1 mutation. This indicates that generalized mitotic recombination between plasmid sequences utilizes functions required for chromosomal recombination in S. cerevisiae.Most laboratory strains of Saccharomyces cerevisiae contain an endogenous plasmid commonly referred to as the 2,u circle (4). Although this plasmid has not yet been shown to carry out an essential function in yeast cells, its properties have made it suitable for studies on yeast DNA replication (15,16,29) and site-specific recombination (5). The origin of replication of this plasmid has also been introduced into a number of yeast cloning vectors (6,24,25) to allow their autonomous, high-copy-number replication.One of the cloning vectors that contains the 2,u circle origin of replication is YEp13 (6). This plasmid also includes the yeast P-isopropyl malate dehydrogenase (LEU2) gene. A derivative of YEp13 has been constructed which contains the yeast arginine permease (CAN!) gene (6). This plasmid (TLC-1) is unstable, because S. cereVisiae strains which contain it can form derivatives which have lost the CANI gene while retaining the LEU2 gene. Such derivatives can be identified because the CANI gene confers dominant sensitivity to the arginine analog canavanine (28), so that loss of the gene from the plasmid results in a canavanine-resistant phenotype in a canl host. This separation of plasmid markers is due to recombination between TLC-1 and the endogenous 2,u circle and involves sequences that are not part of the site-specific recombination system that operates on the inverted repeat sequences. We have used this marker separation to investigate the role of the RAD52 gene product on generalized mitotic recombination between autonomously replicating plasmids in S. cerei'isiae. MATERIALS AND METHODSStrains. S. cerevisiae strains MSW28-1OC(x (leu2-3,2-112 his3-11,3-15 canl-100 iura3-1 trp5-2) and MSW152-lAa (leu2-3,2-112 hisl-7 can!-100 lura3-1 rad52-1) were constructed for this study from strain GRF18a obtained from G. R. Fink.The rad52-1 strain E053-6Doa (rad52-1 arg4-17 ade2-1 lysl-l hisl-7) was supplied by S. Plasmids and probes. Plasmid TLC-1 (6) was obtained from J. Hicks, and pBR322 (3) was obtained from J. Calvo. The LEU2-specifi...

show abstract

Plasmid identification using specific endonucleases

Cited by 95 publications

References 30 publications

Physical Characterization of Ten R Plasmids Obtained from an Outbreak of Nosocomial Klebsiella pneumoniae Infections

Physical Characterization of Ten R Plasmids Obtained from an Outbreak of Nosocomial Klebsiella pneumoniae Infections

Restriction endonuclease characterization of resistant plasmids in Enterobacteriaceae isolated from children in the Sudan

Recombinational instability of a chimeric plasmid in Saccharomyces cerevisiae.

Contact Info

Product

Resources

About