Some properties of nucleic acids extracted with phenol

Segovia, Z.M.Martinez; Sokol, F.; Graves, I. L.; Ackermann, W. Wilbur

doi:10.1016/0005-2787(65)90497-1

Cited by 38 publications

(7 citation statements)

References 6 publications

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“…In experiments in which n-RNA was extracted from the precipitate obtained by treating whole cells with phenol-saturated water (Harris & Watts, 1962) it was found that the rl-RNA was grossly contaminated by polysaccharide. [This finding confirms the observation by Segovia, Sokol, Graves & Ackermann (1965).] When the labelled n-RNA prepared in this way was subjected to sucrose-density-gradient centrifugation, a pellet of material containing a high proportion of the radioactivity was deposited at the bottom of the tube.…”

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confidence: 91%

The origin of the polydispersity in sedimentation patterns of rapidly labelled nuclear ribonucleic acid

Bramwell¹,

Harris²

1967

Biochemical Journal

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1. A study was made of the sedimentation properties of purified preparations of the rapidly labelled RNA in the nucleus and the cytoplasm of the HeLa cell. The sedimentation of the rapidly labelled nuclear RNA was very sensitive to changes in ionic strength and bivalent cation concentration. Under the conditions usually used in sucrose-density-gradient centrifugation the rapidly labelled nuclear RNA showed extreme polydispersity, and much of it sedimented more rapidly than the 28s RNA. At low ionic strength and after removal of Mg(2+), however, the rapidly labelled nuclear RNA sedimented as a single peak at about 16s. The conversion of the polydisperse material into the 16s form did not involve degradation of the RNA, since the effect could be reversed by increasing the ionic strength of the solution. 2. The cytoplasm did not contain any RNA that showed polydisperse sedimentation under the usual conditions of sucrose-density-gradient centrifugation, or that had the same sensitivity as the rapidly labelled nuclear RNA to changes in ionic strength. All the radioactivity in the cytoplasmic RNA sedimented with the 28s, 16s and 4s components over a wide range of physical conditions, but these components did contain a labelled fraction with some of the features of the rapidly labelled nuclear RNA on columns of methylated albumin on kieselguhr. 3. In both nucleus and cytoplasm the RNA detected by ultraviolet absorption could also be converted into a 16s form by removal of bivalent cations at low ionic strength; this effect was again, within certain limits, reversible. The nuclear RNA as a whole was more susceptible to changes in ionic strength than the cytoplasmic RNA. 4. It thus appears that all the RNA in the cell, except the 4s RNA, can be prepared, without degradation, as a single peak sedimenting at about 16s. The relationship of these various 16s components to each other is discussed.

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confidence: 91%

The origin of the polydispersity in sedimentation patterns of rapidly labelled nuclear ribonucleic acid

Bramwell¹,

Harris²

1967

Biochemical Journal

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“…The presence of certain polysaccharides which can appear as satellite bands in CsCI density gradients has been observed in DNA preparations of E. coli (36), photosynthetic bacteria (42), blue-green algae (15), Drosophila (34), and HeLa cells (37). Another possible source of error in the interpretation of these results would derive from the presence of an uncommon base in a portion of the DNA molecule, thus leading to a difference in buoyant density (35).…”

Section: Discussionmentioning

confidence: 99%

Characterization of the Deoxyribonucleic Acid of Various Strains of Halophilic Bacteria

1969

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Bacteria classified as extreme halophiles, in the genera Halobacterium and Halococcus, contain deoxyribonucleic acid (DNA) which displays two components in a CsCI equilibrium density gradient. The base composition of the major DNA component ranges from 66 to 68% guanine plus cytosine (GC), whereas that of the satellite DNA comprising some 11 to 36% of the total, is between 57 and 60% GC. Purification of the bacterial cells in a CsCI density gradient and other more conventional strain purification procedures both indicated that the presence of the satellite DNA component is not a result of mixed cultures.

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“…This fundamental difference in the behavior of dextrans that differed in molecular weight was used in the CsCl density gradient experiments that follow. Band formation by glycogen in CsCl density gradients has also been reported (1,28) and is included in Fig. 11 for reference.…”

Section: Resultsmentioning

confidence: 79%

Streptococcus mutans Dextransucrase: Functioning of Primer Dextran and Endogenous Dextranase in Water-Soluble and Water-Insoluble Glucan Synthesis

Germaine

Harlander²,

Leung³

et al. 1977

Infect Immun

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The extracellular enzyme activities ofStreptococcus mutans 6715 that synthesize glucans from sucrose were concentrated and partially purified by ammonium sulfate precipitation and gel permeation column chromatography. Polyacrylamide gel analysis demonstrated that all of the major proteins precipitated by ammonium sulfate were quantitatively recovered in the high-molecularweight, enzyme-containing aggregates found in the void volume of the gel column. Anion-exchange column chromatography was used to fractionate the aggregates into preparations, a and /3, which produced water-insoluble and water-soluble glucans, respectively. Polyacrylamide gel analysis showed that a and ,3 contained unique proteins and dextransucrase (EC 2.4.1.5) activities. Studies on the time course of glucan synthesis by a demonstrated that this enzyme preparation contained dextranase activity, which partially degraded nascent alcohol-insoluble glucan into alcohol-soluble products that were subsequently reincorporated into insoluble product. The ,3 enzyme preparation contained no detectable dextranase activity. Mixing experiments in the absence of primer dextran demonstrated that the dextranase activity present in a could modify glucan production by /. CsCl density gradient analysis of product glucans demonstrated that exogenous primer dextrans were used as acceptor molecules by both the a and /8 enzyme preparations, and that water-soluble glucans synthesized by /8 could be converted into water-insoluble glucans by a.It is proposed that the structural heterogeneity of the native glucans produced from sucrose by S. mutans is a result of the concerted action of glucan-forming dextransucrases and endohydrolytic dextranase activity. 31). A significant effect of the presence of exogenous dextranase (Spicaria violaceae) during glucan synthesis by S. mutans dextransucrase on the quantity of a-(1--+3) linkages in the product glucans has been reported (7). Thus, the endogenous S. mutans dextranase may con-637 on July 16, 2020 by guest http://iai.asm.org/ Downloaded from cates the position of the bovine serum albumin standard run in an adjacent lane of the gel.

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Some properties of nucleic acids extracted with phenol

Cited by 38 publications

References 6 publications

The origin of the polydispersity in sedimentation patterns of rapidly labelled nuclear ribonucleic acid

The origin of the polydispersity in sedimentation patterns of rapidly labelled nuclear ribonucleic acid

Characterization of the Deoxyribonucleic Acid of Various Strains of Halophilic Bacteria

Streptococcus mutans Dextransucrase: Functioning of Primer Dextran and Endogenous Dextranase in Water-Soluble and Water-Insoluble Glucan Synthesis

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