The Complexity of the High-Molecular-Weight Mammalian Deoxyribonucleic Acid Polymerase Fraction

1973 ◽  
Vol 1 (5) ◽  
pp. 1134-1135 ◽  
Author(s):  
ANDREW M. HOLMES ◽  
IAN P. HESSLEWOOD ◽  
IRVING R. JOHNSTON
1975 ◽  
Vol 149 (1) ◽  
pp. 289-291 ◽  
Author(s):  
G M Blackburn ◽  
P E Taussig

Anthracene becomes covalently bound to high-molecular-weight DNA in mammalian tissue culture as a result of irradiation at 365 nm after the incubation of cells with the hydrocarbon. At high radiation doses, the extent of binding exceeds one hydrocarbon molecule per 103 bases, and is lethal. At low radiation doses, much decreased binding is observed, but a majority of cells remain viable and can be recultured.


1974 ◽  
Vol 2 (5) ◽  
pp. 864-865 ◽  
Author(s):  
ANDREW M. HOLMES ◽  
IAN P. HESSLEWOOD ◽  
WILLIAM F. WAKELING ◽  
IRVING R. JOHNSTON

1974 ◽  
Vol 137 (3) ◽  
pp. 543-546
Author(s):  
G. R. Barker ◽  
P. Hodges

1. Native DNA from two strains of Bacillus subtilis was chromatographed by stepwise elution from MAK (methylated albumin on kieselguhr). 2. Transforming activity was confined to two out of the three main fractions, activity being distributed between the two peaks differently for DNA from the different strains. 3. Fractionation of DNA from both strains on 2% agarose gel gave two components. Approx. 75% of the material was eluted within the void volume of the column. Approx. 25% of the material consisted of degradation products of lower molecular weight. 4. Chromatography on MAK of the material of high molecular weight eluted from agarose gel gave a number of peaks differing in molecular weight, indicating that degradation of the DNA takes place during chromatography on MAK. 5. The distribution of transforming activity among the fractions from MAK suggests that degradation occurs preferentially in certain regions of the DNA.


1975 ◽  
Vol 151 (2) ◽  
pp. 227-238 ◽  
Author(s):  
A G McLennan ◽  
H M Keir

Two DNA polymerases of high molecular weight, pol A (mol.wt. 190 000) and pol B (mol.wt. 240 ooo), have been purified 6300-fold and 1600-fold respectively from an extramitochondrial supernatant of a bleached strain of Euglena gracilis. They have very similar requirements when assayed with an ‘activated’-DNA primer-template [the optimum conditions of pH and ionic (K+ and Mn2+) composition being 7.2, 25 mM and 0.2 mM respectively]. 0.2 mM-Mn2+ was about 1.5-2-fold as effective as 2 mM-Mg2+, owing to substrate activation by deoxyribonucleoside 5′-triphosphates in the presence of Mn2+. Km values for the triphosphates in the absence of activation were about 10(-6)M with Mn2+ and 8 × 10(-6) M with Mg2+ for both enzymes. They were inhibited to the same extent by N-ethylmaleimide, novobiocin and o-phenanthroline, but differed in their chromatographic behaviour on DEAE-cellulose and in their electrophoretic mobilities on polyacrylamide gel. No evidence was found for the existence in these cells of a DNA polymerase of low molecular weight, but there were indications that a third enzyme of high molecular weight might exist.


1972 ◽  
Vol 130 (4) ◽  
pp. 959-964 ◽  
Author(s):  
Leigh A. Burgoyne

During the evaluation of a method of preparing permealysed Ehrlich ascites cells, shortterm labelling experiments were carried out with d[3H]TTP. In the first minute the bulk of the label appeared as low-molecular-weight pieces of DNA. Subsequently the label appeared in DNA of much higher molecular weight. A brief description of the preparation procedure and the properties of the product is provided. Evidence is presented to show that the nucleotide was incorporated directly without intermediate conversion into dTMP or thymidine.


1966 ◽  
Vol 101 (3) ◽  
pp. 647-650 ◽  
Author(s):  
AH Schein

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.


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