Plasmid Curing of <i>Escherichia coli</i> Cells with Ethidium Bromide, Sodium Dodecyl Sulfate and Acridine Orange

The plasmid eliminating abilities of acridine orange, ethidium bromide and sodium dodecyl sulfate were investigated on multi drug resistant Escherichia coli from urinary tract infection specimens. Three different concentrations of each curing agent (Et-Br, SDS and AO) were used. The frequencies of cured cells were 5.55 % (with 50 μg/ml) and 11.76 % (with 75 μg/ml) for acridine orange, 14.29 % (with 100 μg/ml), 21.05 % (with 100 μg/ ml), 17.65 % (with 125 μg/ml) for ethidium bromide and 7.4 % (with 10 % w/v) & 6.67 % (with 10 % w/v) for sodium dodecyl sulfate. However, no cured cells were obtained from 100 μg/ml acridine orange, 75 μg/ml ethidium bromide and 8 and 12 % SDS. Analysis of profiles of wild type and plasmid cured strains by electrophoresis yielded bands of varying sizes for wild type cells, but none were obtained for Et-Br cured cells. Acridine orange treated cells could eliminate only plasmids of 2.7 MDa and another smaller than 2 MDa. Key Words: Plasmid curing; Escherichia coli; Ethidium Bromide; Sodium Dodecyl Sulfate; Acridine Orange. DOI: http://dx.doi.org/10.3329/bjm.v27i1.9165 BJM 2010; 27(1): 28-31

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Sodium Dodecyl Sulfate Hypersensitivity of clpP and clpB Mutants of Escherichia coli

Applied and Environmental Microbiology ◽

10.1128/aem.68.8.4117-4121.2002 ◽

2002 ◽

Vol 68 (8) ◽

pp. 4117-4121 ◽

Cited By ~ 31

Author(s):

Soumitra Rajagopal ◽

Narasimhan Sudarsan ◽

Kenneth W. Nickerson

Keyword(s):

Escherichia Coli ◽

Sodium Dodecyl Sulfate ◽

Uronic Acid ◽

Sodium Dodecyl ◽

Immunoblot Analysis ◽

Wild Type ◽

E Coli ◽

Dodecyl Sulfate ◽

Shock Proteins ◽

Western Immunoblot Analysis

ABSTRACT We studied the hypersensitivity of clpP and clpB mutants of Escherichia coli to sodium dodecyl sulfate (SDS). Both wild-type E. coli MC4100 and lon mutants grew in the presence of 10% SDS, whereas isogenic clpP and clpB single mutants could not grow above 0.5% SDS and clpA and clpX single mutants could not grow above 5.0% SDS. For wild-type E. coli, cellular ClpP levels as determined by Western immunoblot analysis increased ca. sixfold as the levels of added SDS increased from 0 to 2%. Capsular colanic acid, measured as uronic acid, increased ca. sixfold as the levels of added SDS increased from 2 to 10%. Based on these findings, 3 of the 19 previously identified SDS shock proteins (M. Adamowicz, P. M. Kelley, and K. W. Nickerson, J. Bacteriol. 173:229-233, 1991) are tentatively identified as ClpP, ClpX, and ClpB.

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Identification of the glpT -Encoded sn -Glycerol-3-Phosphate Permease of Escherichia coli , an Oligomeric Integral Membrane Protein

Journal of Bacteriology ◽

10.1128/jb.152.3.1008-1021.1982 ◽

1982 ◽

Vol 152 (3) ◽

pp. 1008-1021

Author(s):

Timothy J. Larson ◽

Günter Schumacher ◽

Winfried Boos

Keyword(s):

Escherichia Coli ◽

Molecular Weight ◽

Sodium Dodecyl Sulfate ◽

Gene Product ◽

Active Transport ◽

Sodium Dodecyl ◽

Apparent Molecular Weight ◽

Wild Type ◽

Dodecyl Sulfate ◽

Active Transport System

A collection of hybrid plasmids carrying either the wild-type or mutated glpT gene was generated in vitro and used to characterize the glpT -dependent active transport system for sn -glycerol-3-phosphate in Escherichia coli K-12. Restriction endonuclease analysis and recloning of DNA fragments localized glpT to a 3-kilobase pair Pst I- Hpa I segment of DNA. Comparison of DNA carrying glpT-lacZ fusions with DNA carrying intact glpT allowed determination of the direction of transcription. Through characterization of the proteins synthesized by strains harboring hybrid plasmids carrying amber, missense, or deletion mutations in glpT , it was shown that glpT is a promoter-proximal gene in an operon consisting of at least two genes. The gene product of glpT , the sn -glycerol-3-phosphate permease, was found associated with the inner membrane. It could be solubilized by treatment with sodium dodecyl sulfate at 50°C. Its molecular weight, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was dependent upon sample treatment before electrophoresis. The apparent molecular weight was 44,000 when membrane fractions were heated to 50°C; subsequent treatment at 95°C modified the protein such that it migrated faster (apparent molecular weight = 33,000). Several missense mutations in glpT were negatively dominant over wild-type glpT , indicating that the active form of the permease is multimeric. A gene (named glpQ ) promoter distal to glpT codes for a periplasmic protein. This protein had previously been named GLPT protein to indicate its relationship to the glpT gene. The present report demonstrates that it is not the gene product of glpT and is not required for active transport of sn -glycerol-3-phosphate.

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Discrimination of live and dead cells of Escherichia coli using propidium monoazide after sodium dodecyl sulfate treatment

Food Control ◽

10.1016/j.foodcont.2016.06.022 ◽

2017 ◽

Vol 71 ◽

pp. 79-82 ◽

Cited By ~ 14

Author(s):

Hajime Takahashi ◽

Yun Gao ◽

Satoko Miya ◽

Takashi Kuda ◽

Bon Kimura

Keyword(s):

Escherichia Coli ◽

Sodium Dodecyl Sulfate ◽

Sodium Dodecyl ◽

Propidium Monoazide ◽

Dodecyl Sulfate

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Ubiquitin immunoreactivity shows several proteins varying with development and sporulation in the basidiomycete Coprinus cinereus

Biochemistry and Cell Biology ◽

10.1139/o90-150 ◽

1990 ◽

Vol 68 (7-8) ◽

pp. 1019-1025 ◽

Cited By ~ 5

Author(s):

Tosaku Kanda ◽

Nobuharu Tanaka ◽

Tsuneo Takemaru

Keyword(s):

Sodium Dodecyl Sulfate ◽

Sodium Dodecyl ◽

Light Period ◽

Coprinus Cinereus ◽

Polyacrylamide Gels ◽

Wild Type ◽

Stages Of Development ◽

Crude Extracts ◽

Dodecyl Sulfate ◽

Molecular Masses

Crude extracts of mycelia and basidiocarp primordia in the basidiomycete Coprinus cinereus were resolved on sodium dodecyl sulfate – polyacrylamide gels, and ubiquitin and several proteins were detected by immunoblotting with anti-ubiquitin antibody. The molecular masses of the proteins detected were 30 900, 28 600, 27 800, 26 300, 22 500, and 15 400 daltons, respectively. Relative levels of ubiquitin and the ubiquitin-immunoreactive proteins were measured in different stages of development. The levels of ubiquitin and most of the ubiquitin-immunoreactive proteins in basidiocarp primordium formation increased and in basidiocarp maturation decreased in cap and upper stipe, while in lower stipe became high except for the 27 800 dalton protein and ubiquitin. During sporulation, ubiquitin and all the ubiquitin-immunoreactive proteins tended to decrease in the cap of the young wild-type basidiocarp. The levels of 30 900 and 15 400 dalton proteins increased transiently at 6–10 h after the beginning of the last light period, while ubiquitin decreased markedly. No correlation was observed between changes in levels of the ubiquitin-immunoreactive proteins and the blocked stages in sporulation-deficient mutants.Key words: ubiquitin, development, sporulation, Coprinus.

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