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

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.

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 Ligation and supercoiling of plasmids containing single strand interruptions

2021 ◽  
Author(s):  
Felipe A. Calil ◽  
Christopher D. Putnam ◽  
Richard D. Kolodner
Keyword(s):  
Gel Electrophoresis ◽  
Simple Procedure ◽  
Dna Gyrase ◽  
Single Strand ◽  
Dna Ligase ◽  
Hydroxyl Groups ◽  
Agarose Gel ◽  
Agarose Gel Electrophoresis ◽  
Circular Dna ◽  
T4 Dna Ligase

Abstract We have developed a simple procedure for determining if a plasmid DNA contains single strand interruptions that can be sealed by DNA ligase and thus contain single strand interruptions with 5'-phosphate and 3'-hydroxyl groups. In this procedure, plasmid DNAs with ligatable nicks were ligated by T4 DNA ligase, supercoiled by E. coli DNA gyrase and analyzed by agarose gel electrophoresis. Our results show that after sealing nicked circular DNA with DNA ligase, supercoiling by DNA gyrase produces a rapidly migrating DNA species that can easily be distinguished from nicked circular DNA by agarose gel electrophoresis.

Plasmid DNA Topology Assayed by Two-Dimensional Agarose Gel Electrophoresis

2013 ◽  
pp. 121-132 ◽  
Author(s):  
Jorge B. Schvartzman ◽  
María-Luisa Martínez-Robles ◽  
Pablo Hernández ◽  
Dora B. Krimer
Keyword(s):  
Plasmid Dna ◽  
Gel Electrophoresis ◽  
Dna Topology ◽  
Agarose Gel ◽  
Two Dimensional ◽  
Agarose Gel Electrophoresis

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.

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