scholarly journals Kinetic complexity of chloroplastal deoxyribonucleic acid and mitochondrial deoxyribonucleic acid from higher plants

1969 ◽  
Vol 112 (5) ◽  
pp. 777-786 ◽  
Author(s):  
Richard Wells ◽  
Max Birnstiel
Keyword(s):  
Molecular Weight ◽  
Mitochondrial Dna ◽  
Gaussian Distribution ◽  
Deoxyribonucleic Acid ◽  
Nuclear Dna ◽  
Higher Plants ◽  
Buoyant Density ◽  
Cell Fractionation ◽  
Caesium Chloride ◽  
Cellular Dna

1. Chloroplasts and mitochondria were isolated by aqueous and non-aqueous cell-fractionation techniques. In a variety of higher plants the mitochondrial DNA bands in a caesium chloride gradient at 1·706g.cm.−3, whereas chloroplastal DNA has a buoyant density of 1·697g.cm.−3. 2. In total cellular DNA of moderate molecular weight, the chloroplastal DNA is found within the Gaussian distribution of the nuclear DNA and is not resolved as a satellite. 3. Both chloroplastal DNA and mitochondrial DNA from lettuce renature rapidly. 4. The kinetic complexity of mitochondrial DNA is > 108 daltons. 5. Chloroplastal DNA is made up from fast and slow renaturing sequences with kinetic complexities of 3×106 and 1·2×108 daltons respectively. 6. From the discrepancy between analytical and kinetic complexity it is concluded that chloroplastal DNA is extensively reiterated.

scholarly journals Mitochondrial deoxyribonucleic acid from Tetrahymena pyriformis and its kinetic complexity

1970 ◽  
Vol 116 (5) ◽  
pp. 811-817 ◽  
Author(s):  
R. A. Flavell ◽  
I. G. Jones
Keyword(s):  
Mitochondrial Dna ◽  
Melting Temperature ◽  
Density Gradient ◽  
Deoxyribonucleic Acid ◽  
Nuclear Dna ◽  
Buoyant Density ◽  
Tetrahymena Pyriformis ◽  
Base Pairing ◽  
Caesium Chloride ◽  
Kinetics Of

1. Mitochondrial DNA from Tetrahymena pyriformis strain T has a buoyant density (ρ) of 1.685 compared with ρ1.688 for whole cell DNA. Mitochondrial preparations from T. pyriformis strain W show an enrichment of a light satellite (ρ1.686), although this is not obtained free from nuclear DNA (ρ1.692). 2. T. pyriformis mitochondrial DNA renatures rapidly and the kinetics of this process indicate a complexity of approx. 3×107 daltons. 3. The base-pairing in the renaturation product is of a precise nature, since the ‘melting’ temperature (80.5°C) is indistinguishable from that of the native DNA (80.5°C). 4. Centrifugation of mitochondrial DNA in an alkaline caesium chloride density gradient gives two bands, implying the separation of the complementary strands.

scholarly journals MINOR COMPONENTS OF THE DNA OF PHYSARUM POLYCEPHALUM

1969 ◽  
Vol 40 (2) ◽  
pp. 484-496 ◽  
Author(s):  
Charles E. Holt ◽  
Elizabeth G. Gurney
Keyword(s):  
Cell Cycle ◽  
Physarum Polycephalum ◽  
Nuclear Dna ◽  
Buoyant Density ◽  
Cell Fractionation ◽  
Minor Components ◽  
Variable Level ◽  
The Mean ◽  
S Period ◽  
Dna Components

DNA metabolism in the slime mold Physarum polycephalum was studied by centrifugation in CsCl of lysates of cultures labeled with radioactive thymidine at various times in the cell cycle. During the G2 (premitotic) phase of the cell cycle, two components of the DNA are labeled. One component is lighter (buoyant density 1.686 g/cc) than the mean of the principal DNA (1.700 g/cc), and one is heavier (approximately 1.706 g/cc). The labeled light DNA was identified chemically by its denaturability, its susceptibility to DNase, and the recovery of its radioactivity in thymine. Cell fractionation studies showed that the heavy and the principal DNA components are located in the nucleus and that the light DNA is in the cytoplasm. The light DNA comprises approximately 10% of the DNA. About ⅓–½ of the light DNA is synthesized during the S period, and the remainder is synthesized throughout G2 (there is no G1 in Physarum). The light DNA is metabolically stable. A low, variable level of incorporation of radioactive thymidine into the principal, nuclear DNA component was observed during G2.

DNA Synthesis in Isolated Yeast Mitochondria

1974 ◽  
Vol 52 (11) ◽  
pp. 941-949 ◽  
Author(s):  
L. Zeman ◽  
C. V. Lusena
Keyword(s):  
Molecular Weight ◽  
Mitochondrial Dna ◽  
Base Composition ◽  
Nuclear Dna ◽  
Sedimentation Velocity ◽  
Yeast Saccharomyces Cerevisiae ◽  
Denatured State ◽  
Saccharomyces Cerevisiae Mitochondria

Isolated yeast (Saccharomyces cerevisiae) mitochondria incorporate radioactive precursors into mitochondrial DNA. This in vitro labelled DNA was characterized by isopycnic and sedimentation velocity centrifugation both in the native and denatured state. The profiles of isopycnic CsCl gradients obtained by centrifugation in a fixed-angle rotor are skewed toward high density. The skew is neither due to the presence of in vitro labelled nuclear DNA nor due to random breaks in mitochondrial DNA which would reveal, then, its heterogeneity in base composition. The in vitro labelled DNA is reproducibly recovered as a class of molecules sedimenting at about 5–8 S, indicating a molecular weight of 1 × 105 – 4 × 105 daltons, while the smallest in vivo labelled fragments sediment at about 13–14 S, corresponding to 1.6 × 106 – 2.0 × 106 daltons. After denaturation, the in vitro labelled DNA molecules sediment at about 2–5 S, corresponding to a single-strand molecular weight of 1 × 104 – 7 × 104 daltons, which is about one hundred times less than the observed size of the denatured in vivo labelled molecules.

scholarly journals NUCLEAR DNA AND CYTOPLASMIC DNA FROM TISSUES OF HIGHER PLANTS

1965 ◽  
Vol 27 (3) ◽  
pp. 451-457 ◽  
Author(s):  
Yasuo Hotta ◽  
Alix Bassel ◽  
Herbert Stern
Keyword(s):  
Inorganic Phosphate ◽  
Nuclear Dna ◽  
Higher Plants ◽  
Soluble Fraction ◽  
Buoyant Density ◽  
Wheat Roots ◽  
Cytoplasmic Dna ◽  
Sucrose Medium

Young wheat roots were labeled with 32P-inorganic phosphate. Following the labeling period, roots were homogenized in a sucrose medium and fractionated into nuclei, cytoplasmic particles (including proplastids and mitochondria), and a soluble fraction containing most of the microsomes. DNA prepared from the particles had a higher buoyant density than that from the nuclei and showed a marked loss in total label if the roots were exposed to non-radioactive medium for 48 hours prior to fractionation of the cells.

scholarly journals The deoxyribonucleic acid of Micrococcus radiodurans

1966 ◽  
Vol 101 (3) ◽  
pp. 647-650 ◽  
Author(s):  
AH Schein
Keyword(s):  
Electron Microscopy ◽  
Molecular Weight ◽  
High Molecular Weight ◽  
Deoxyribonucleic Acid ◽  
High Molecular Weight Dna ◽  
Caesium Chloride

The DNA of Micrococcus radiodurans was prepared by three methods. Although the recovery of DNA varied considerably, the percentage molar base ratios of the DNA from the three preparations were essentially the same: guanine, 33+/-2; adenine, 18+/-1; cytosine, 33+/-2; thymine, 17+/-1. Base compositions calculated from T(m) values and from density in caesium chloride gradients also yielded guanine+cytosine contents of 66 and 68% of total bases respectively. No unusual bases were observed. The S(20,w) values were characteristic of high-molecular-weight DNA. Electron microscopy showed the purified DNA in long strands; occasionally these were coiled.

scholarly journals Some Physicochemical Properties of Erysimum Latent Virus

1982 ◽  
Vol 35 (1) ◽  
pp. 5
Author(s):  
Keith H Gough ◽  
Glenn G Lilley ◽  
Dharma D Shukla ◽  
Frank Woods
Keyword(s):  
Molecular Weight ◽  
Buoyant Density ◽  
Latent Virus ◽  
Sedimentation Equilibrium ◽  
Protein Subunit ◽  
Molecular Weights ◽  
Caesium Chloride ◽  
Rna Content ◽  
Velocity Diffusion ◽  
Phosphorus Determination

Sedimentation velocity, diffusion coefficient and sedimentation equilibrium measurements gave a molecular weight of 5 �90 x 106 for the intact Erysimum latent virus. The molecular weight of the empty shell was estimated to be 3�92 X 106 and the protein subunit to be 21 600. The RNA content calculated from the molecular weights of the full and empty particles is 33 %, in agreement with that estimated from the buoyant density in caesium chloride. However, a direct phosphorus determination gave an RNA content of only 28 %.

scholarly journals The use of equilibrium-density-gradient methods for the preparation and characterization of blood-group-specific glycoproteins

1970 ◽  
Vol 117 (5) ◽  
pp. 879-891 ◽  
Author(s):  
J. M. Creeth ◽  
M. A. Denborough
Keyword(s):  
Molecular Weight ◽  
Density Gradient ◽  
Blood Group ◽  
Buoyant Density ◽  
Gradient Methods ◽  
Apparent Molecular Weight ◽  
Sedimentation Equilibrium ◽  
Equilibrium Density ◽  
Caesium Chloride ◽  
Selective Solvation

1. The method of sedimentation equilibrium in a gradient of caesium chloride has been applied to the preparation of blood-group-specific glycoproteins from human ovarian-cyst fluids: it is shown that virtually complete separation from contaminating protein is easily accomplished in a single step. 2. The glycoproteins isolated in this way have been characterized by analytical density-gradient experiments in both caesium chloride and caesium sulphate and values of the buoyant density, selective solvation and apparent molecular weight have been obtained. 3. In some cases, materials prepared from the same cysts by solvent extraction methods have also been characterized in these terms. 4. The selective solvation values are about 0.1 and 0.5g of water/g of glycoprotein in caesium chloride and caesium sulphate respectively. 5. The apparent molecular-weight values are much lower than the weight-average molecular weights, and it is shown that the origin of the discrepancy is heterogeneity in density of the glycoproteins. 6. Some sources of error in the interpretation of density-gradient schlieren patterns are examined.

Chloroplast deoxyribonucleic acid Isolation: purity achieved without nuclease digestion

1981 ◽  
Vol 59 (11-12) ◽  
pp. 911-915 ◽  
Author(s):  
Patsy R. Rhodes ◽  
S. D. Kung
Keyword(s):  
Chloroplast Dna ◽  
Liquid Nitrogen ◽  
Deoxyribonucleic Acid ◽  
Nuclear Dna ◽  
Buoyant Density ◽  
Silica Sol ◽  
Plant Tissues ◽  
Nuclease Digestion ◽  
Density Analysis ◽  
Digestion Step

Chloroplasts, obtained from plant tissues homogenized in liquid nitrogen, were freed of nuclei on silica sol gradients. Buoyant density analysis of denatured–renatured DNA and the clarity of restriction fragment patterns demonstrate the purity of these preparations. In this manner, chloroplast DNA free of substantial nuclear DNA contamination was obtained from several plant species without the use of a deoxyribonuclease digestion step.

Nondestructive sampling of mitochondrial DNA from voles (Microtus)

1987 ◽  
Vol 65 (1) ◽  
pp. 175-180 ◽  
Author(s):  
Yves Plante ◽  
Peter T. Boag ◽  
Bradley N. White
Keyword(s):  
Mitochondrial Dna ◽  
Population Structure ◽  
Social Interactions ◽  
Nuclear Dna ◽  
Microtus Pennsylvanicus ◽  
Dna Polymorphisms ◽  
Biochemical Taxonomy ◽  
Meadow Voles ◽  
Blood Samples ◽  
Cellular Dna

We present two techniques for sampling mitochondrial DNA (mtDNA) without killing individual voles. Total cellular DNA was extracted from small blood samples (100–250 μL) and tail segments (2 cm long) collected from meadow voles (Microtus pennsylvanicus). Restriction fragment patterns produced by the restriction endonucleases HindIII, BamH1, and EcoR1 after hybridization with a probe of nick-translated mtDNA compared well with standard mtDNA assays. Both techniques can be used in the field, and should prove useful in biochemical taxonomy as well as in investigations of population structure, dispersal, and social interactions on both micro- and macro-geographic scales. The Southern blots produced by these procedures have the added advantage of being reuseable, so that nuclear DNA polymorphisms can be examined with appropriate probes.

Genome Sizes of Chloroplast and Mitochondrial DNA's in Higher Plants

1972 ◽  
Vol 30 ◽  
pp. 190-191
Author(s):  
Richard Kolodner ◽  
K.K. Tewari
Keyword(s):  
Molecular Weight ◽  
Higher Plants ◽  
Buoyant Density ◽  
Molecular Size ◽  
Mt Dna ◽  
A Value ◽  
Ct Dna ◽  
Kinetic Class

Chloroplast (ct-) and mitochondrial (mt-) DNA's from pea, spinach, bean, and lettuce leaves have been studied for their molecular size and conformation. The ct-DNA's from these plants were found to band at a density of 1.698 ± 0.001 g/cm3 and denatured with a Tm of 84° ± 0.5°C. The corresponding mt-DNA's had a buoyant density of 1.706 ± 0.001 g/cm3 and denatured with a Tm of 88° ± 0.5°C. Both ct- and mt-DNA from these plants were found to have a homogeneous melting pattern. The sheared and denatured ctand mt-DNA's from these plants were found to renature as a single kinetic class with no indication of repeating sequences. The molecular weight of the ct-DNA from the higher plants studied has been found to be 95 ± 5 x 106 by renaturation rates assuming a value of 106 x 106 for T4 DNA.

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