Retention of ribosomal ribonucleic acids in agarose gels

Petrović, Silvana; Petrović, Jelena; Novaković, Milka B.

doi:10.1016/0005-2787(73)90161-5

Cited by 19 publications

(6 citation statements)

References 21 publications

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“…3). This is in agreement with our observations on the separation of DNA from highmolecular-weight RNA by agarose chromatography (Petrovic et al, 1973).…”

Section: Resultssupporting

confidence: 93%

“…Recent findings on the retention behaviour of polynucleotides in agarose gels (Petrovic et al, 1971(Petrovic et al, , 1973Morris et al, 1972) indicated the possibility of improvements in the purification of double-stranded RNA species by agarose chromatography at an increased ionic strength. As shown in the present paper, it was possible to define conditions that result in selective retention or exclusion from the gel of nearly all major types of nucleic acids found in a Vol.…”

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confidence: 99%

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Fractionation of nucleic acids from Penicillium chrysogenum and associated ribonucleic acid viruses by selective exclusion and retention in agarose gels

Petrović¹,

Šumonja²,

Vasiljević³

1974

Biochemical Journal

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Double-stranded nucleic acids from a strain of Penicillium chrysogenum containing RNA viruses were isolated by agarose-gel filtration, and separated into DNA and double-stranded RNA fractions by agarose-gel chromatography in 2.5m-NaCl. The DNA fraction contained less than 1% alkali-labile polynucleotides, and sedimented homogeneously at 8-10S in alkaline sucrose gradients. In CsCl gradients it tended to band in the density region of 1.66-1.72g/ml. It had a ;melting' temperature (T(m)) of 75 degrees C in 0.015m-NaCl-0.0015m-trisodium citrate, corresponding to 51.5mol% of G+C. The double-stranded RNA fraction did not contain detectable DNA. It could not band in CsCl up to a density of 1.78g/ml, and mainly consisted of a 14-15S RNA species with a T(m) of 88.5 degrees C in the above solvent, and a G+C content of 49.3 mol%.

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“…3). This is in agreement with our observations on the separation of DNA from highmolecular-weight RNA by agarose chromatography (Petrovic et al, 1973).…”

Section: Resultssupporting

confidence: 93%

mentioning

confidence: 99%

Fractionation of nucleic acids from Penicillium chrysogenum and associated ribonucleic acid viruses by selective exclusion and retention in agarose gels

Petrović¹,

Šumonja²,

Vasiljević³

1974

Biochemical Journal

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“…However, this is not the case for nucleic acids, as observed in this and in an earlier publication (Petrovió et al, 1973(Petrovió et al, , 1974aWoo et al, 1974). The unusual chromatographic behavior of RNA may be explained by the major role of secondary structure, as suggested by Petrovic et al (1973Petrovic et al ( , 1974a. High ionic strength, which stabilizes secondary structure, increases retention of RNAs on Sepharose.…”

Section: Discussionmentioning

confidence: 99%

“…High ionic strength, which stabilizes secondary structure, increases retention of RNAs on Sepharose. The ionic strength at which RNAs are eluted from the column depends on the GC content and the molecular weight of the RNA and tends to decrease as molecular weight increases (Petrovic et al, 1973). This property of the RNA and Sepharose 4B to interact provides a useful method to separate RNAs with small differences in molecular weight but which differ in base composition.…”

Section: Discussionmentioning

confidence: 99%

Resolution of ribonucleic acids by Sepharose 4B column chromatography

Zeichner¹,

Stern²

1977

Biochemistry

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Ribonucleic acids were resolved by molecular sieve chromatography on columns of Sepharose 4B. The elution positions of messenger ribonucleic acids were determined by detection of polyadenosine tracts and by support of protein synthesis in a messenger-dependent cell-free system. The elution position of other ribonucleic acid species from the Sepharose 4B was determined by formamide-sucrose density gradient centrifugation. Resolution of ribonucleic acids by this column was not dependent on molecular weight but rather on other properties such as secondary structure or the presence of poly(adenylic acid). The elution profiles of ribonucleic acids on cross-linked Sepharose 4B differed markely from those on conventional Sepharose and appeared to depend on molecular size alone. There was diminished resolution of high molecular weight ribonucleic acids on such columns.

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“…Sci. USA 74 (1977) 4553 The mating efficiency of a cross was calculated according to Chiang et al (25) For DNA preparation for CsCl buoyant density analytical centrifugation, the proteinase K-digested cell lysate was adjusted to 1 M in NaClO4 and extracted with a 24:1 (vol/vol) mixture of chloroform/isoamyl alcohol (26), and the aqueous phase of this extraction was passed through an agarose column (Bio-Gel A, 50-m; Bio-Rad Laboratories, Richmond, CA) equilibrated with 2 M NaCl (27). The DNA, detected by absorbance at 254 nm, eluted with the void volume and was pooled and dialyzed against 0.15 M NaCl/0.015 M sodium citrate.…”

Section: Methodsmentioning

confidence: 99%

Perturbation of chloroplast DNA amounts and chloroplast gene transmission in Chlamydomonas reinhardtii by 5-fluorodeoxyuridine

Wurtz

Boynton²,

Gillham³

1977

Proc. Natl. Acad. Sci. U.S.A.

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5-Fluorodeoxyuridine selectively decreases the rate of chloroplast DNA replication in Chlamydomonas resulting after several generations of growth in equilibrium levels as low as one-seventh of normal. When the maternal parent is treated prior to mating, the decrease of chloroplast DNA alppears to perturb the normal maternal transmission of chloroplast genes, dramatically increasing the proportion of exceptional zygotes transmitting chloroplast genes from the paternal parent. Uniparental inheritance in the isogamous green alga Chlamydomonas reinhardtil was discovered more than 20 years ago by Sager (1). Much information about this genetic system, now thought to reside in chloroplast DNA, has been obtained since (2-6). Multiple copies of this DNA are believed by most workers to be present in the single chloroplast of C. reinhardtfl because the absolute amount of chloroplast DNA per cell is 25-50 times the molecular weight of an individual chloroplast DNA molecule. This is true whether molecular weight is determined by kinetic complexity (7)(8)(9), by size measurements of intact molecules observed in the electron microscope (10), or by summation of the molecular weights of EcoRl restriction fragments (11, 12). However, the mechanisms responsible for the uniparental inheritance of chloroplast DNA and chloroplast markers as well as the number of genetic copies transmitted to meiotic progeny are still in dispute (2-4, 6, 13-15).Sager has proposed a mechanism of uniparental inheritance modeled after the restriction-modification systems in bacteria (13,14). In most zygotes from a cross, paternal chloroplast DNA is destroyed by a restriction enzyme while maternal chloroplast DNA is modified by methylation and hence is protected from restriction. All progeny receive two copies of the chloroplast genome. In contrast, our laboratory (3, 4) has proposed a multicopy model with certain analogies to a phage cross. The input of maternal and paternal chloroplast genomes is variable and the output of genomes from the two parents depends upon their relative success in competing for attachment sites in the single zygote chloroplast that results from fusion of the two gametic chloroplasts (16). Normally, these sites are occupied by maternal chloroplast genomes.To test directly which of the two foregoing models is correct we have used the thymidine analogue 5-fluorodeoxyuridine (FdUrd) to perturb the amount of chloroplast DNA and then observed the effect of this analogue on the transmission of chloroplast genes. Our rationale was as follows. In Chlamydomonas, exogenous radioactive thymidine labels chloroplast, but not nuclear, DNA in vegetative cells and cells undergoing gametogenesis (17,18) MATERIALS AND METHODS C. reinhardtll wild-type strain 137c+ (GB-125) and antibiotic-resistant strains carrying the chloroplast markers spr-u-1-6-2, mt-(GB-146, paternal parent) and er-u-37, sr-u-2-60, mt + (GB-148, maternal parent) isolated from this strain (3,4) were grown either phototrophically in high-salt medium bubbled with 5% CO...

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Retention of ribosomal ribonucleic acids in agarose gels

Cited by 19 publications

References 21 publications

Fractionation of nucleic acids from Penicillium chrysogenum and associated ribonucleic acid viruses by selective exclusion and retention in agarose gels

Fractionation of nucleic acids from Penicillium chrysogenum and associated ribonucleic acid viruses by selective exclusion and retention in agarose gels

Resolution of ribonucleic acids by Sepharose 4B column chromatography

Perturbation of chloroplast DNA amounts and chloroplast gene transmission in Chlamydomonas reinhardtii by 5-fluorodeoxyuridine

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