Paper Chromatography of Several Classes of Compounds: Correlated R<sub>f</sub> Values in a Variety of Solvent Systems

Fink, Kay; Cline, Richard E.; Fink, Robert M.

doi:10.1021/ac60196a036

Cited by 184 publications

(38 citation statements)

References 12 publications

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“…DNA was analyzed by the methods of Fink et ai. (9,10) and its buoyant density in CsC1 determined according to Meselson et al (11), as described in the previous publication (1). Although D29 DNA was characterized previously, it was included in the experiments reported here for comparative purposes; the values obtained agree, within the limits of experimental error, with those previously reported (1).…”

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confidence: 75%

Further Studies of Infectious Dna Extracted From Mycobacteriophages

Sellers

Tokunaga

1966

The Journal of Experimental Medicine

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In the previous publication (1) it was reported that deoxyribonucleic acid (DNA) extracted from mycobacteriophage D29 could infect its host, Mycobacterium smegmatis 607. No alteration in the structural integrity of the bacteria such as the production of protoplasts was required; caltures susceptible to intact phage could also be infected with phage D N A .This investigation has been extended to include three additional mycobacteriophages. I n the experiments reported here, D N A extracted from D4 and D32 but not that from D28, was found to be infectious. A t t e m p t s to extend the host range of intact phage with phage D N A were not successful. Materials and MethodsThe source and preparation of reagents, enzymes, buffers, and chromatography procedures were the same as those described in the preceding publication (1). One additional buffer used in this study, Versonate-potassium trichioroacetate (V-KTCA), Nutrional Biochemicals Company, Cleveland, Ohio, was prepared according to Weil (2). Versonate buffer was composed of 0.2 g disodium Versonate, NaCI 8.0 g, KC1 0.2 g, Na2HPO, 1.15 g, KHdaO4 0.02 g, distilled water to 1000 ml. Potassium trichioroacetate: 0.15 ~, pH 7.6. The final V-KTCA buffer was made up of 1.5 parts Versonate and 2.5 parts potassium trichioroacetate. Compounds not previously described (1), will be noted in the text.Pkages.--D28, D29, D32, and D4 were obtained originally from Dr. Seymour Froman.They were isolated by soil-enrichment techniques and except for D4, are active against certain strains of both virulent and saprophytic acid-fast bacilli; D4 is active against Mycobacterium smegmatis 607 (3, 4). Methods for the production of high-titered lysates and for phage assay have been described (5).Baaeria.--Mycobaaerium smegmatis ATCC 607 (propagating host for D4 and for D29), Mycobacterium sp. ATCC9033 (propagating host for D32), and Mycobacterium sp. ATCC 9626 (propagating host for D28) were employed. These bacteria will be referred to in the text by their ATCC numbers. Two additional species of Mycobacterium were employed in hostrange experiments. They were obtained from Dr. Froman and, according to him, one is M.phlei (F 89) and the other M. smegmalis (F 87). The latter can be distinguished from 607 by its phage susceptibility pattern. These bacteria will be referred to in the text as F 89 and as F 87. Stock cultures were maintained, and log-phase cultures prepared as described (5).

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supporting

confidence: 75%

Further Studies of Infectious Dna Extracted From Mycobacteriophages

Sellers

Tokunaga

1966

The Journal of Experimental Medicine

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“…The nucleosides were separated by two-dimensional paper chromatography, by the basic technique described previously (26,27). An amount of extract equivalent to 2.2 A U at 260 nm was applied as a small spot on Whatman No.…”

Section: Methodsmentioning

confidence: 99%

Hereditary Hemolytic Anemia with Human Erythrocyte Pyrimidine 5′-Nucleotidase Deficiency

Valentine¹,

Fink²,

Paglia³

et al. 1974

J. Clin. Invest.

258

141

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A B S T R A C T A severe deficiency of a red cell pyrimidine 5'-nucleotidase was found to be associated with hereditary hemolytic anemia in four members of three kindreds. The syndrome was characterized by marked increases above normal in red cell basophilic stippling, total nucleotides, and GSH and by a fairly severe deficiency of ribosephosphate pyrophosphokinase (EC 2.7. 6.1.). Patient erythrocytes uniquely contained large amounts of pyrimidine 5'-ribonucleotides. In earlier studies, these were erroneously considered to be adenosine phosphates, since all previous investigations of the nucleotides of human red cells and reticulocytes have shown 97% or more to contain adenine. Total nucleotides in patient cells were present in amounts 3-6 times greater than normal, and approximately 80% contained pyrimidine. The ultraviolet spectral curves of deproteinized red cell extracts exhibited a shift in maximum absorbance from the usual 256-257 nm to approximately 266-270 nm, and absorbance at 250, 270, 280, and 290 nm, expressed as a ratio of that at 260 nm, differed greatly from normal. The spectral characteristics of extracts provide the basis of a readily performed screening procedure, which does not require enzyme assay. The nucleotidase activity in deficient red cells assayed less than 14%, and usually less than 10%, of normal and much less in terms of reticulocyte-rich blood, where it was consistently found to be increased. The enzyme has a pH optimum of 7.5-8.0, is inhibited by EDTA, and does not utilize purine 5'-ribonucleotides or 8-glycerophosphate as substrates. While comparatively few family members have been available thus far for study, initial data are compatible with an autosomal, recessive

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“…The procedures for separation of the nucleosides by two dimensional paper chromatography and the subsequent estimation of their concentrations from ultraviolet absorbance spectrophotometric data have been described previously (1,24,25). …”

Section: Methodsmentioning

confidence: 99%

Lead poisoning: association with hemolytic anemia, basophilic stippling, erythrocyte pyrimidine 5'-nucleotidase deficiency, and intraerythrocytic accumulation of pyrimidines.

Valentine¹,

Paglia²,

Fink³

et al. 1976

J. Clin. Invest.

105

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Paper Chromatography of Several Classes of Compounds: Correlated R_f Values in a Variety of Solvent Systems

Cited by 184 publications

References 12 publications

Further Studies of Infectious Dna Extracted From Mycobacteriophages

Further Studies of Infectious Dna Extracted From Mycobacteriophages

Hereditary Hemolytic Anemia with Human Erythrocyte Pyrimidine 5′-Nucleotidase Deficiency

Lead poisoning: association with hemolytic anemia, basophilic stippling, erythrocyte pyrimidine 5'-nucleotidase deficiency, and intraerythrocytic accumulation of pyrimidines.

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Paper Chromatography of Several Classes of Compounds: Correlated Rf Values in a Variety of Solvent Systems

Cited by 184 publications

References 12 publications

Further Studies of Infectious Dna Extracted From Mycobacteriophages

Further Studies of Infectious Dna Extracted From Mycobacteriophages

Hereditary Hemolytic Anemia with Human Erythrocyte Pyrimidine 5′-Nucleotidase Deficiency

Lead poisoning: association with hemolytic anemia, basophilic stippling, erythrocyte pyrimidine 5'-nucleotidase deficiency, and intraerythrocytic accumulation of pyrimidines.

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Paper Chromatography of Several Classes of Compounds: Correlated R_f Values in a Variety of Solvent Systems