Characterization and transformation of plasmid pAA-1 found in an antarctic cryptoendolithic bacterium

1996 ◽  
Vol 42 (6) ◽  
pp. 571-576 ◽  
Author(s):  
Christian G. Gliesche ◽  
Marina Jendrach ◽  
Klaus Peissl ◽  
Jörg Siebert ◽  
Peter Hirsch
Keyword(s):  
Plasmid Dna ◽  
Gel Electrophoresis ◽  
Natural Transformation ◽  
Restriction Map ◽  
Agarose Gel ◽  
Dry Valleys ◽  
Agarose Gel Electrophoresis ◽  
Bacterial Isolates ◽  
E Coli ◽  
Natural Genetic Transformation

Sixty-three bacterial isolates from antarctic sandstone samples (Linnaeus Terrace, Asgard Range, McMurdo Dry Valleys) were screened for the presence of plasmid DNA. Twenty-seven percent of all the isolates (mainly Gram-positives) harbored one or more plasmids of low molecular mass (1.1–16.3 kb). Strain AA-341 contained plasmid pAA-1 (2.9 kb), as demonstrated by agarose gel electrophoresis and restriction endonuclease digests. This plasmid conferred resistance to chromium and ampicillin. It was not conjugative, but it could be transferred by electroporation to chromium- and ampicillin-sensitive strains AA-330, AA-338, and E. coli HB101. A restriction map of pAA-1 was constructed with HindIII, SalI, ScaI, AvaII, EcoRI, PvuII, BamHI, and DraI. Electrotransfer of this plasmid from E. coli HB101(pAA-1) to strain AA-330 was demonstrated. By natural genetic transformation, plasmid pAA-1 could be transferred into the sensitive strain AA-334, which thereby became resistant to chromium and ampicillin. The importance of such processes for the colonization of stressed environments is discussed.Key words: Antarctica, cryptoendolithic bacteria, plasmids, resistance to chromium, natural transformation.

scholarly journals Bacterial Isolates from Bivalve Clams (Galatea paradoxa, Born 1778): Occurence, Multi-drug Resistance, Location of Antibiotic Resistance Marker and Plasmid Profiles

2020 ◽  
pp. 35-46
Author(s):  
M. U. Okon ◽  
C. U. Inyang ◽  
O. J. Akinjogunla
Keyword(s):  
Drug Resistance ◽  
Antibiotic Resistance ◽  
Plasmid Dna ◽  
Gel Electrophoresis ◽  
Multi Drug Resistance ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Bacterial Isolates ◽  
Salmonella Spp ◽  
Galatea Paradoxa

The occurrence of bacterial isolates in Galatea paradoxa (Born 1778) was determined using standard bacteriological method. The multi-drug resistance, location of antibiotic markers, plasmid DNA extraction and electrophoresis was determined by disc diffusion, acridine orange, TENS alkaline lysis and 0.8% agarose gel electrophoresis, respectively. Of the 63 bacterial isolates from G. paradoxa, Staphylococcus aureus and Streptococcus pyogenes had the highest and lowest percentage of occurrence with 40.0% and 5.0%, respectively. Escherichia coli was 25.0%, Pseudomonas aeruginosa (17.5%), Enterococcus spp and Salmonella spp (15.0%) each, Bacillus subtilis (12.5%), Klebsiella pneumoniae and Enterococcus faecalis (10.0%) each while Vibrio cholerae was (7.5%). The results showed Streptomycin and Ciprofloxacin as the most effective antibiotics against bacterial isolates from G. paradoxa. Bacillus subtilis and P. aeruginosa displayed 100% sensitivity to Streptomycin; Salmonella spp and E. faecalis were 100% sensitive to Augmentin. V. cholerae and S. pyogenes showed 100% resistance to Penicillin and Rifampicin, respectively. Of the 63 bacterial isolates, 43 (68.3%) were multidrug resistant (MDR) isolates, of which S. aureus and E. coli had the widest multiple antibiotic resistance (MAR) indices ranging from 0.3 to 0.8, while S. pyogenes had the least MAR ≤ 0.5. Of the 43 MDR bacterial isolates, 16.3%, 23.3% and 60.5% had their entire antibiotic resistance encoded on plasmid, chromosome and both plasmid and chromosome, respectively. The agarose gel electrophoresis showed that MDR bacterial isolates from G. paradoxa had plasmid DNA with molecular weights ranging from 23.1 to 31.5kb. This study has showed that G. paradoxa harboured bacteria which could pose serious health risks and G. paradoxa should be adequately cooked before consumption.

Simple procedure for distinguishing CCC, OC, and L forms of plasmid DNA by agarose gel electrophoresis

Plasmid ◽  
1981 ◽  
Vol 5 (3) ◽  
pp. 371-373 ◽  
Author(s):  
G. Hintermann ◽  
H.-M. Fischer ◽  
R. Crameri ◽  
R. Hütter
Keyword(s):  
Plasmid Dna ◽  
Gel Electrophoresis ◽  
Simple Procedure ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis

scholarly journals Plasmid DNA replication and topology as visualized by two-dimensional agarose gel electrophoresis

Plasmid ◽  
2010 ◽  
Vol 63 (1) ◽  
pp. 1-10 ◽  
Author(s):  
J.B. Schvartzman ◽  
M.L. Martínez-Robles ◽  
P. Hernández ◽  
D.B. Krimer
Keyword(s):  
Dna Replication ◽  
Plasmid Dna ◽  
Gel Electrophoresis ◽  
Agarose Gel ◽  
Two Dimensional ◽  
Agarose Gel Electrophoresis ◽  
And Topology

A Bacteriocin Produced by Pediococcus Species Associated with a 5.5-Megadalton Plasmid

1988 ◽  
Vol 51 (1) ◽  
pp. 29-31 ◽  
Author(s):  
DALLAS G. HOOVER ◽  
PATRICIA M. WALSH ◽  
KAREN M. KOLAETIS ◽  
MARY M. DALY
Keyword(s):  
Listeria Monocytogenes ◽  
Plasmid Dna ◽  
Gel Electrophoresis ◽  
Leuconostoc Mesenteroides ◽  
Agarose Gel ◽  
Pediococcus Acidilactici ◽  
Plasmid Curing ◽  
Agarose Gel Electrophoresis ◽  
Bacteriocin Activity ◽  
Zones Of Inhibition

Agarose gel electrophoresis of plasmid DNA and plasmid-curing experiments suggested that bacteriocin activity was harbored on a plasmid of approximately 5.5 megadaltons (Mdal) in Pediococcus acidilactici PO2, B5627 and PC, and Pediococcus pentosaceus MC-03. Bacteria that were sensitive to the bacteriocin included other pediococci, Leuconostoc mesenteroides, Streptococcus faecalis and Listeria monocytogenes. Three of the five serotypes of L. monocytogenes examined were inhibited by a variant lacking the 5.5 Mdal plasmid; however, when the plasmid was present, zones of inhibition doubled in size.

Transformation of Pseudomonas syringae with nonconjugative R plasmids

1981 ◽  
Vol 27 (8) ◽  
pp. 759-765 ◽  
Author(s):  
Dennis C. Gross ◽  
Anne K. Vidaver
Keyword(s):  
Molecular Weight ◽  
Pseudomonas Syringae ◽  
Plasmid Dna ◽  
Gel Electrophoresis ◽  
Recipient Cell ◽  
Heat Pulse ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Colony Forming Units ◽  
R Plasmids

Transformation of Pseudomonas syringae strains with plasmid DNA occurs at a frequency of 1 × 10−3 to 4 × 10−9 per recipient cell, depending on the strain, plasmid, and conditions for transformation. R plasmids used successfully in transformation were pR0161 (26 × 106 molecular weight) and RSF1010 (5.5 × 106 molecular weight). Transformation involved growing the recipient cells to approximately 8 × 108 colony-forming units per millilitre in 50 mL of a nutrient broth. After washes with a 150 mM CaCl2 – 10% (v/v) glycerol mixture, cells were concentrated 20-fold and resuspended in this solution. The cells then were incubated with purified plasmid DNA for 1 h prior to a heat pulse at 45 °C for 2 min. Transformants were selected by antibiotic resistance and plasmid presence was verified by agarose gel electrophoresis. With plasmid pCG131 (34 × 106 molecular weight; putatively associated with syringomycin production), transformation of syringomycin-negative P. syringae strains that contained no detectable plasmid or were cured of pCG131 was unsuccessful, whether the plasmid was used alone or in combination with either pR0161 or RSF1010.

scholarly journals Curing of Escherichia coli Antibiotic Resistance Plasmids by Aspirin

2014 ◽  
Vol 8 (1) ◽  
pp. 30-35
Author(s):  
Ibtesam H. Al Musawi ◽  
Ali A. Al Zaag
Keyword(s):  
Escherichia Coli ◽  
Gel Electrophoresis ◽  
Agarose Gel ◽  
Large Plasmid ◽  
Agarose Gel Electrophoresis ◽  
E Coli ◽  
Resistance Plasmids ◽  
Minimum Number

An isolate of Escherichia coli (E. coli) was isolated from urine sample due to person infected with urinary tract infection (UTI).The isolate was resistant to the following antibiotics: Ampicillin, Cotrimoxazole, Chloramphenicol and Tetracycline. Agarose gel electrophoresis of its plasmids content has revealed the presence of single large plasmid and two small plasmids bands. The large plasmid was conjugative and contained the resistance genes for four antibiotics. In vitro curing of this plasmid was achieved by treatment with salicylic acid (aspirin) with 150,200,250 and 300 µg/ml as indicated by the elimination of resistance, also by absence of large plasmid band following agarose gel electrophoresis. In vivo curing was conducted using New Zealand rabbits. UTI was induced by bacterial inoculation via urethral catheterization. The E. coli from urine samples of the rabbits, the count of which was proportional to the type of treatment. Minimum number of colonies was associated with group treated with metheprim was aspirin 300mg/kg daily dosage. This result may indicate that the side effect of metheprim was in its maximum with aspirin. Survivor bacteria may indicate incomplete exposure to the drug.

scholarly journals Genetic study of Salmonella spp. Producting Betalatimase (ESBLs)

2010 ◽  
Vol 7 (1) ◽  
pp. 254-260
Author(s):  
Baghdad Science Journal
Keyword(s):  
Genetic Study ◽  
Gel Electrophoresis ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Biochemical Tests ◽  
Salmonella Spp ◽  
Small Plasmid ◽  
E Coli ◽  
Extended Spectrum ◽  
Synergy Test

Ten isolates were collected from different clinical sources from laboratory in medicine century . These isolates were belonging to the genus Salmonella depending on morphological and biochemical tests . The antibiotic scussptibility tests against 10 antibiotics were examined , and it was found that the 60% isolates have multiple resistant to antibiotic ,(70%) of isolates were resistant to ampicillin,(50%) were resistant to augmentin ,(40%) were resistant to ceftriaxone ,(20%) were resistant to cefotaxime and (10%) were resistant to ciprofloxacin and tetracycline while all isolates showed sensitivity to piperacillin, imipenem, amikacin and erythromycin .The ability of Salmonela isolates to produce ?-lactamase enzymes were tested using iodometric method , and the results showed that all isolates produced this enzyme.The ability of these isolates to produce Extended-Spectrum ?-lactamase (ESBLs)were also determined by double disc synergy test , only five isolates produced these enzyme. Agarose gel electrophoresis showed that Salmonella isolates ?-lactamase producer have two small plasmid bands . Transformation experiments revealed that these plasmids were capable to transform E. coli MM294, an observation which indicates the ability of these plasmids to show their expression in more than one host.

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