A rapid method for analysis of ligand binding to deoxyribonucleic acid and soluble nucleoproteins using streptomycin: application to steroid receptor ligands. 1983

T C Spelsberg

A method is described which allows the rapid analysis of the binding of practically any molecules to DNA or to protein--DNA complexes (termed nucleoacidic protein or NAP). The antibiotic streptomycin sulfate, a soluble aminoglycoside, is used to precipitate the DNA after the ligand binding. Comparison of different sources and commercial batches of the antibiotic is described. Optimal conditions for precipitating DNA or NAP and the application of this method to the binding of the chick oviduct progesterone receptor to soluble NAP are described. The streptomycin method can be used with DNA molecules whose size ranges from 750 base pairs to greater than 50 000 base pairs. The method works with a DNA or NAP from a variety of sources, including synthetic homo- or heteropolymers. The precipitation of DNA or NAP by streptomycin occurs rapidly and has minimal effects on the steroid receptor complex or binding of the steroid receptor to DNA or NAP. The requirements and limitations of the method as well as the optimal conditions for binding of the progesterone receptor to DNA or NAP are described.

UI MeSH Term Description Entries
D008722 Methods A series of steps taken in order to conduct research. Techniques,Methodological Studies,Methodological Study,Procedures,Studies, Methodological,Study, Methodological,Method,Procedure,Technique
D008970 Molecular Weight The sum of the weight of all the atoms in a molecule. Molecular Weights,Weight, Molecular,Weights, Molecular
D009698 Nucleoproteins Proteins conjugated with nucleic acids. Nucleoprotein
D010057 Oviducts Ducts that serve exclusively for the passage of eggs from the ovaries to the exterior of the body. In non-mammals, they are termed oviducts. In mammals, they are highly specialized and known as FALLOPIAN TUBES. Oviduct
D011980 Receptors, Progesterone Specific proteins found in or on cells of progesterone target tissues that specifically combine with progesterone. The cytosol progesterone-receptor complex then associates with the nucleic acids to initiate protein synthesis. There are two kinds of progesterone receptors, A and B. Both are induced by estrogen and have short half-lives. Progesterone Receptors,Progestin Receptor,Progestin Receptors,Receptor, Progesterone,Receptors, Progestin,Progesterone Receptor,Receptor, Progestin
D002621 Chemistry A basic science concerned with the composition, structure, and properties of matter; and the reactions that occur between substances and the associated energy exchange.
D002645 Chickens Common name for the species Gallus gallus, the domestic fowl, in the family Phasianidae, order GALLIFORMES. It is descended from the red jungle fowl of SOUTHEAST ASIA. Gallus gallus,Gallus domesticus,Gallus gallus domesticus,Chicken
D004247 DNA A deoxyribonucleotide polymer that is the primary genetic material of all cells. Eukaryotic and prokaryotic organisms normally contain DNA in a double-stranded state, yet several important biological processes transiently involve single-stranded regions. DNA, which consists of a polysugar-phosphate backbone possessing projections of purines (adenine and guanine) and pyrimidines (thymine and cytosine), forms a double helix that is held together by hydrogen bonds between these purines and pyrimidines (adenine to thymine and guanine to cytosine). DNA, Double-Stranded,Deoxyribonucleic Acid,ds-DNA,DNA, Double Stranded,Double-Stranded DNA,ds DNA
D005260 Female Females
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia

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