Subject(s)
Erythrocyte Membrane/drug effects , Fishes/blood , Receptors, Adrenergic, beta/drug effects , Adenylyl Cyclases/metabolism , Adrenergic beta-Agonists/pharmacology , Animals , Bucladesine/pharmacology , Cell Membrane Permeability , Colforsin/pharmacology , Enzyme Activation , Erythrocyte Membrane/enzymology , Erythrocyte Membrane/physiology , Ion Transport , Norepinephrine/pharmacology , Polychlorinated Biphenyls/pharmacology , Receptors, Adrenergic, beta/physiology , Sodium-Potassium-Exchanging ATPase/antagonists & inhibitorsABSTRACT
On the basis of the reaction of dibromoethylacetate with adenine and cytosine at the uncoiled regions of DNA it was possible to fix these molten regions (the degree of denaturation was about 16%) for rat liver mitochondrial DNA. Modification of all accessible adenine and cytosine residues in 0.14 M Na-acetate, pH 5.65, was completed at 70-80 degrees C in 15-20 min. Using the computer orientation for the set of fixed BamHI-fragments and the linear molecules of full length the denaturation map of mtDNA was constructed, the GC-content of the molten regions was about 28%. The dibromoethylacetate is a perspective agent for screening DNA with a low content of destabilizing regions (for instance, DNA of malignant cells).
Subject(s)
DNA, Mitochondrial/genetics , Ethylene Dibromide/pharmacology , Hydrocarbons, Brominated/pharmacology , Mitochondria, Liver/metabolism , Adenine , Animals , Cytosine , DNA Restriction Enzymes , Deoxyribonuclease BamHI , Nucleic Acid Conformation , Nucleic Acid Denaturation , RatsABSTRACT
The nature of intramolecular heterogeneity of mtDNA in the liver of white rats has been studied. The peculiarities of the melting curve, and the possibility of DNA fractionation of nucleotide compounds with hydroxylapatite (HA) column chromatography has shown the presence of sequences differing in the mean nucleotide content. A section of about 350 pairs in size repeated four times was found in the reassociation of most thermolabile fraction with a mean composition of 28% GC. These sections are well seen on the denaturation map of the recorded molecules formed in the range of temperature transition 'helix-coil'. The distance between the centers of fusible sections (in percentage of total length) is 32.5, 32, 14.0 and 21.5.
