Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “plasmid characterization”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 145 records · Page 8Linked to original sources
PubMed1978

Characterization of plasmids in bacteriocin producing strains of Streptococcus faecium.

Four bacteriocin (enterocin) producing strains of Streptococcus faecium were examined for extrachromosomal DNA. Three strains proved to harbor minicircular plasmid DNA of uniform length as evidenced by CsCl ethidium bromide density centrifugation and electron microscopy. Molecular weight determination obtained through contour length measurements showed 2.4 X 10(6) dalton for S. faecium 25, 3.4 X 10(6) dalton for S. faecium 3 and 3.8 X 10(6) dalton for S. faecium 59. Elimination experiments for bacteriocin production with intercalating dyes proved sterile. The role of these plasmids remains for the time being unknown.

Anti-Bacterial Agents↗
PubMed1991

Partial characterization of plasmids from rabbit isolates of Pasteurella multocida.

Plasmids have not been reported for isolates of Pasteurella multocida from rabbits. We assayed 28 isolates of rabbit P. multocida for plasmids and sought to determine whether or not plasmid presence correlated with clinical or pathologic findings, serotype, toxin production, possession of pili, or biochemical characteristics. Fourteen isolates bore a single 1.6 Md (covalently closed circular form in 0.7% agarose gels) plasmid. An additional isolate had two plasmids which migrated as a closely-spaced doublet, centered around 1.6 Md. Eleven isolates appeared to have identical plasmids, according to Hae III and Hinf I digests. The apparent linear size of this common plasmid in 2% agarose gels was 2.1 Md, as calculated from the sums of the sizes of Hae III or Hinf I digestion fragments. Linearization of the common plasmid with Msp I produced an apparent size of 2.5 Md in 0.7% agarose gels. No correlations between presence of the common plasmid and somatic serotype, toxigenicity, presence of pili, antimicrobial resistance, selected biochemical characteristics, anatomic site from which the bacteria were cultured, or disease status of the host were found.

Animals↗
PubMed1984

[Epidemiological surveillance of beta-lactamase-producing gonococci. II--Characterization of plasmids of 66 strains isolated in France (May 1979-March 1983)].

Penicillinase producing Neisseria gonorrhoeae PPNG (66 strains have been examined for the presence of plasmids coding for bêta-lactamase. The 3.2 Mdal plasmid was present in 27/66 (41%) of strains. The 4.5 Mdal plasmid was present in 30/66 (45%) of strains. The association 2.6 Mdal 4.5 Mdal plasmids were only present in 9/66 (13.6%) of strains. The cryptic plasmid of 2.6 Mdal was always present.

Drug Resistance, Microbial↗
PubMed1982

Occurrence and characterization of plasmids in field isolates of Bordetella bronchiseptica.

Twenty-seven isolates of Bordetella bronchiseptica were examined for the presence of plasmid DNA. Eleven of the isolates contained plasmids. An attempt was made to correlate the presence of plasmids with resistance to a number of antibiotics or heavy metals or with production of bacteriocins. Eight of the isolates contained a plasmid of approximately 35 megadaltons molecular weight; these isolates were all resistant to 1 mM mercuric chloride. There were no other correlations between plasmid content and any of the phenotypes tested. The plasmids conferring resistance to mercuric chloride were transferred by conjugation to Escherichia coli K-12. The plasmid-bearing isolates were then compared with their isogenic plasmidless parent for resistance to a number of antibiotics. Isolates acquiring the plasmids demonstrated increased resistance to streptomycin and ampicillin, as well as to mercuric chloride. The 8 plasmids were also analyzed by restriction endonuclease digestion.

Animals↗
PubMed1995

Intratracheal gene delivery to the mouse airway: characterization of plasmid DNA expression and pharmacokinetics.

Intratracheal administration of plasmid DNA resulted in gene expression in mouse airways in the absence of any enhancing agent. Administration of plasmid DNA encoding the chloramphenicol acetyltransferase gene (CAT) in sterile water lead to CAT transgene expression that peaked between 1 and 3 days and was detected up to 28 days after DNA administration. Transgene expression was independent of mouse gender, age and strain. Levels of expression from DNA in various isotonic solutions did not differ from levels obtained with DNA administered in water, suggesting that transfection is not dependent on damage to airway cells caused by a hypo-osmotic delivery vehicle. Pharmacokinetic studies using radiolabeled plasmid DNA showed that DNA was rapidly degraded, while higher levels of radioactivity were retained for longer duration following administration of cationic liposome-DNA complexes in the airway. Southern blot and PCR analysis confirmed that DNA complexed with DOTMA-DOPE was retained in the airways for a longer period. However, cationic liposomes DOTMA-DOPE (1:1) or DOTAP complexed with DNA, did not enhance expression over DNA alone. These results suggest that 'naked' plasmid DNA should be included as a control in all studies on intratracheal gene delivery using nonviral systems.

Animals↗
PubMedDate not supplied

[Characterization of plasmids which mediate resistance to multiple antibiotics in gram-negative bacteria of nosocomial origin].

BACKGROUND: The genetic and molecular mechanisms involved in antimicrobial resistance of 10 strains of gramnegative bacilli (1 Serratia marcescens; 2 Escherichia coli; 1 Proteus mirabilis; 4 Klebsiella pneumoniae; 1 Enterobacter cloacae y 1 Alcaligenes faecalis), isolated from adult patients with nosocomial pulmonary infection at the in-patient facilities of the University Hospital of Los Andes, Mérida, Venezuela, have been studied. METHODS: The antimicrobial susceptibility was determined by minimum inhibitory concentrations using the dilution method in agar. The study of extrachromosomal genes was carried out by conjugation, bacterial infection with the bacteriophage M13 and curing of plasmid by acridine orange. The plasmids were isolated by alkaline lysis and analysis of restriction endonuclease digestion was carried out separately using the enzymes EcoRI and HindIII. A DNA probe, derived from the region which encodes the TEM-1 beta-lactamase of the plasmid pBR322 was used for dot-blot hybridization tests. RESULTS: All of the gramnegative bacilli showed resistance to ampicillin, carbenicillin and cephalothin (> 128 micrograms/ml) and 3 strains also showed resistance to gentamicin (> 64 micrograms/ml). Genetic and molecular procedures showed the presence of conjugative plasmids of approximately 54 kb in all the 10 strains. The restriction patterns obtained by using EcoRI and HindIII indicated common DNA fragments in most of the plasmids studied. The dot-blot hybridization tests confirmed homology between the plasmids and the DNA probe used (TEM-1 beta-lactamase). CONCLUSIONS: In this study, the gramnegative bacteria of nosocomial origin harbored self-transferable plasmids of approximately 54 kb, which mediate resistance to gentamicin and encode a beta-lactamase of the TEM group.

Adult↗
PubMed1998

Manufacturing and quality control of plasmid-based gene expression systems.

DNA plasmid-based gene expression systems are being widely investigated for the potential treatment of genetic and acquired disease and for DNA-based vaccination. A number of human clinical trials are in progress using plasmid-based drugs. The regulatory framework that has been applied to biologicals such as recombinant DNA-derived proteins has proven to be generally applicable for regulating plasmid-based drugs as well. This was recently emphasized by the inclusion of therapeutic DNA plasmid products in the U.S. Food and Drug Administration's list of well-characterized biotechnology products. Present techniques for manufacturing and characterizing plasmids have been adapted from large-scale protein purification and from traditional molecular biology. Production of multi-gram quantities of plasmid, at purities of 95% or more, is currently possible, but further development of both manufacturing and analytical techniques is required. This review describes the approaches and methods currently used to manufacture and characterize DNA plasmids for pharmaceutical use, as well as recent changes in the regulatory environment that will impact future development and marketing of plasmids as human drugs.

Journal Article↗
PubMed1990

Plasmid-mediated beta-lactam resistance in pathogenic gram-negative bacteria isolated in south India.

A field study was undertaken at the Christian Medical College Hospital in Vellore, South India in 1984. Two hundred and eighty-four clinical isolates of enterobacteria were collected from patients with significant bacteriuria and their sensitivities to a series of beta-lactam antibiotics were determined. There was a very high incidence of beta-lactam resistance (ampicillin resistance greater than 80% and cephaloridine resistance greater than 65%) amongst these isolates. There was significant resistance to the combination of ampicillin and clavulanic acid but few of the isolates were resistant to cefuroxime or ceftazidime. Seventy seven per cent of the Escherichia coli isolates were resistant to ampicillin and 57% were resistant to cephaloridine. Most of the resistant Esch. coli isolates carried resistance genes for both ampicillin and cephaloridine which were transferable, either by direct conjugation or by mobilization with the X+ plasmid. Characterization of the R-plasmids revealed considerable diversity. Although the distribution of plasmid-mediated beta-lactamases was broadly similar to those found in other surveys, three new beta-lactamases were identified and atypically some of the Esch. coli isolates produced PSE beta-lactamases.

Ampicillin↗
PubMed2000

Exogenous isolation of antibiotic resistance plasmids from piggery manure slurries reveals a high prevalence and diversity of IncQ-like plasmids.

Antibiotic resistance plasmids were exogenously isolated in biparental matings with piggery manure bacteria as plasmid donors in Escherichia coli CV601 and Pseudomonas putida UWC1 recipients. Surprisingly, IncQ-like plasmids were detected by dot blot hybridization with an IncQ oriV probe in several P. putida UWC1 transconjugants. The capture of IncQ-like plasmids in biparental matings indicates not only their high prevalence in manure slurries but also the presence of efficiently mobilizing plasmids. In order to elucidate unusual hybridization data (weak or no hybridization with IncQ repB or IncQ oriT probes) four IncQ-like plasmids (pIE1107, pIE1115, pIE1120, and pIE1130), each representing a different EcoRV restriction pattern, were selected for a more thorough plasmid characterization after transfer into E. coli K-12 strain DH5alpha by transformation. The characterization of the IncQ-like plasmids revealed an astonishingly high diversity with regard to phenotypic and genotypic properties. Four different multiple antibiotic resistance patterns were found to be conferred by the IncQ-like plasmids. The plasmids could be mobilized by the RP4 derivative pTH10 into Acinetobacter sp., Ralstonia eutropha, Agrobacterium tumefaciens, and P. putida, but they showed diverse patterns of stability under nonselective growth conditions in different host backgrounds. Incompatibility testing and PCR analysis clearly revealed at least two different types of IncQ-like plasmids. PCR amplification of total DNA extracted directly from different manure samples and other environments indicated the prevalence of both types of IncQ plasmids in manure, sewage, and farm soil. These findings suggest that IncQ plasmids play an important role in disseminating antibiotic resistance genes.

Animals↗
PubMed1991

Plasmids in diatom species.

We have discovered plasmids in 5 of 18 diatom species surveyed. In several species, more than one type of plasmid is present. Several of the plasmids show similarity by hybridization previously characterized plasmids in Cylindrotheca fusiformis (J. D. Jacobs et al., unpublished data). Additionally, there is similarity between the plasmids found in C. fusiformis and chloroplast DNA in three diatom species. These results add to the evidence that the plasmids have features of mobile genetic elements.

Cloning, Molecular↗
PubMed1993

The giant linear plasmid pHG207 from Rhodococcus sp. encoding hydrogen autotrophy: characterization of the plasmid and its termini.

As described previously, in Rhodococcus sp. (formerly Nocardia opaca) strains MR11 and MR22, the ability to grow as an aerobic hydrogen bacterium (the Aut character) is located on giant conjugative linear plasmids--on pHG201 (270 kb) in strain MR11 and on pHG205 (280 kb) in strain MR22. In an autotrophic transconjugant originating from MR22 a smaller plasmid, pHG207 (225 kb), was detected and shown to be a recombination product of the wild-type plasmids pHG204 and pHG205. A donor carrying pHG207 as the sole plasmid transferred the Aut marker at a 1000-fold frequency compared to the wild-type plasmid pHG205. Analysis of the plasmid ends revealed that plasmid pHG207 carries proteins at both ends; the proteins are linked to the 5' ends of the strands. The cloned end fragments of about 2 kb were sequenced and found to contain highly homologous sequences within the terminal 583 bp (left end part) and 560 bp (right end part). Several potential reading frames were detected, but database searching gave no indication about possible functions.

Bacterial Proteins↗
PubMed1985

Characterization of R-plasmids coding for ampicillin resistance in Salmonella typhi-murium.

Eight Salmonella typhi-murium strains coding resistance to ampicillin were chosen from 38 strains isolated in different counties of Hungary in 1981, and their plasmids were characterized by agarose gel electrophoresis. Incompatibility groups and molecular weights of transferable R-plasmids coding resistance to ampicillin were determined and restriction enzyme analyses were done. The studies showed that among R-plasmids coding for ampicillin resistance in S. typhi-murium strains IncI alpha group plasmids with a molecular weight of 66 Mdal were dominant in Hungary. According to their cleavage patterns, the examined plasmids formed two groups. Both types contained some fragments identical in size, and they are supposed to be connected evolutionarily.

Ampicillin↗
PubMed1979

Isolation and characterization of recombinant DNA plasmids carrying Drosophila tRNA genes.

Recombinant plasmids carrying Drosophila melanogaster tRNA genes were constructed by ligation of HindIII-cleaved Drosophila DNA to HindIII cut pBR322 DNA. 90 clones were isolated that contained genes for one or more of eleven tRNAs. 43 of the plasmids were characterized by a number of methods: restriction nuclease digestion; agarose gel electrophoresis; hybridization with individual, purified, 125I-labelled Drosophila tRNA molecules and in situ hybridization to Drosophila chromosomes. The results show that several different tRNA genes have been isolated which code for single, specific isoacceptors. The DNAs from 8 plasmids each hybridize to single sites on Drosophila polytene chromosomes. In addition, the data show examples of two different plasmids hybridizing to different loci coding for the same tRNA; this means that we have isolated representatives of tRNA genes which map at widely separated points on the Drosophila genome.

Animals↗