Polyacrylamide Slab Gel Electrophoresis of Soluble Proteins for Studies of Bacterial Floras

Moore, W. E. C.; Hash, D. E.; Holdeman, Lillian V.; Cato, Elizabeth P.

doi:10.1128/aem.39.4.900-907.1980

Cited by 144 publications

(64 citation statements)

References 17 publications

(22 reference statements)

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“…Although Moore et al (1985) did not uniformly sample progressive periodontal lesions to represent disease, they also found that W. recta and B. gracilis were related to diseased sites. Moore et al (1985) found W. curva and C. concisus were related to diseased sites. In this investigation, we did not identify W. curva from deep periodontal sites and few C. concisus strains were isolated.…”

Section: Discussionmentioning

confidence: 92%

“…It is of interest that B. gracilis is isolated with increasing frequency from deep extra-oral infections (Johnson, Reinhardt & Finegold 1985). Although Moore et al (1985) did not uniformly sample progressive periodontal lesions to represent disease, they also found that W. recta and B. gracilis were related to diseased sites. Moore et al (1985) found W. curva and C. concisus were related to diseased sites.…”

Section: Discussionmentioning

confidence: 99%

“…Attempts were made to differentiate this *Present address; Department of Periodontics, University of Texas Health Center, San Antonio, Texas, U.S.A, set of small oral asaccharolytic rods using sensitivities to inhibitors (dyes, indicators, antibiotics) (Tanner et al , 1984a and serological reactions , Tanner & Ebersole 1982. More recently, polyacrylamide gel eiectrophoresis has been used to differentiate (Moore et al 1980) and identify these species (Tanner 1986).…”

Section: Introductionmentioning

confidence: 99%

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Wolinella recta, Campylobacter concisus, Bacteroides gracilis, and Eikeneila corrodens from periodontal lesions

Tanner¹,

Dzink²,

Ebersole

et al. 1987

J of Periodontal Research

110

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The fastidious gram negative asaccharolytic rods Wolinella recta, Campylobacter concisus, Bacteroides gracilis, and Eikeneila corrodens have been isolated from lesions of advanced periodontal disease. In this investigation, three methods were compared for their ability to identify fresh isolates of these species: 1) biochemical tests and sensitivities to dyes and other inhibitors, 2) serological reactions, and 3) sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS‐PAGE). SDS‐PAGE of total cell proteins appeared to be the most sensitive and accurate method to identify isolates, although many strains were successfully identified using serology. Biochemical tests including sensitivities to inhibitors were the least reliable. W. recta, B. gracilis, and E. corrodens were found more frequently and in higher numbers in progressive (active) disease sites of many of the subjects. E. corrodens was also found to be significantly elevated in inactive sites of certain subjects. After successful treatment with Widman flap surgery and systemically administered tetracycline, the proportions of W. recta and E. corrodens were reduced, while those of C. concisus were elevated.

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Section: Discussionmentioning

confidence: 92%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Wolinella recta, Campylobacter concisus, Bacteroides gracilis, and Eikeneila corrodens from periodontal lesions

Tanner¹,

Dzink²,

Ebersole

et al. 1987

J of Periodontal Research

110

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“…Also Moore etal. (13) and Tanner (29) have differentiated oral bacteria by PAGE of cellular proteins.…”

Section: Discussionmentioning

confidence: 99%

Outer membrane proteins of Actinobacillus actinomycetemcomitans and Haemophilus aphrophilus studied by SDS‐PAGE and immunoblotting

Bolstad¹,

Kristoffersen²,

Olsen³

et al. 1990

Oral Microbiology and Immunology

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This investigation characterized and compared outer membrane proteins (OMP) of the closely related Actinobacillus actinomycetemcomitans and Haemophilus aphrophilus by means of SDS-PAGE patterns and reactions on immunoblots with rabbit antiserum against A. actinomycetemcomitans FDC Y4. Reactions with serum from a patient with Papillon Lefévre syndrome (PLS), from whom periodontal wild strains of A. actinomycetemcomitans had been isolated, were also studied. OMP were purified with selective solubilization from lyophilized cells of 10 wild and 4 reference strains of A. actinomycetemcomitans and 4 reference strains of H. aphrophilus. OMP profiles from wild and reference strains of A. actinomycetemcomitans were similar while those from A. actinomycetemcomitans and H. aphrophilus differed. The most prominent difference was absence of a heat modifiable protein in H. aphrophilus strains. Immunoblotting revealed strong common antigens in most strains, including a heat modifiable protein with mol wt 34 kDa, as well as a 29 kDa and a 16.5 kDa protein. Treatment with pronase and sodium periodate confirmed the protein nature of the major OMP antigens.

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“…Gram-positive cocci are among the most difficult bacteria to disrupt by physical methods (Dobrogosz 1981). Shaking staphylococci with glass beads potentially gives small scale, highly variable extraction (Moore et a!. 1980).…”

mentioning

confidence: 99%

A procedure for urease and protein extraction from staphylococci

Sissons

Hancock

Perinpanayagam

et al. 1989

Journal of Applied Bacteriology

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Staphylococcal cell protein and urease can be solubilized after growth in Todd-Hewitt broth supplemented with 0.5% yeast extract by extraction for 18-24 h in phosphate buffer, pH 7.0. In general 20% (but up to 100%) of the urease present in the original cells could be solubilized. Less protein was solubilized. Species examined included coagulase-negative staphylococci, Staphylococcus intermedius and Staph. aureus. Extracts of Staph. epidermidis prepared by this procedure gave electrophoretic urease and protein patterns similar to those prepared by sonication. The procedure was simple and minimized handling of the cells.

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Polyacrylamide Slab Gel Electrophoresis of Soluble Proteins for Studies of Bacterial Floras

Cited by 144 publications

References 17 publications

Wolinella recta, Campylobacter concisus, Bacteroides gracilis, and Eikeneila corrodens from periodontal lesions

Wolinella recta, Campylobacter concisus, Bacteroides gracilis, and Eikeneila corrodens from periodontal lesions

Outer membrane proteins of Actinobacillus actinomycetemcomitans and Haemophilus aphrophilus studied by SDS‐PAGE and immunoblotting

A procedure for urease and protein extraction from staphylococci

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