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P M Silverman 《Archives of biochemistry and biophysics》1973,157(1):234-242
The number of RNA sites bound by the Qβ RNA polymerase and the affinity of the enzyme for different RNA sites was measured by equilibrium partition of enzyme and enzyme-nucleic acid complexes between two liquid polymer phases. At 0 °C and in the absence of other components required for RNA synthesis the enzyme bound to many regions of Qβ RNA, f2 RNA, 16S rRNA, double-stranded RNA, and circular DNA. Under conditions of enzyme excess, the maximum numbers of enzyme molecules bound per molecule of Qβ RNA, f2 RNA, and 16S rRNA were 32, 26, and 12, respectively. The enzyme bound to most, if not all, of the Qβ RNA sites with the same affinity. Nevertheless, the association constant for enzyme binding to Qβ RNA was more than 10-fold greater than for binding to f2 RNA over a wide range of salt concentrations. 相似文献
996.
Purification and analysis of murine 2-5A-dependent RNase 总被引:6,自引:0,他引:6
R H Silverman D D Jung N L Nolan-Sorden C W Dieffenbach V P Kedar D N SenGupta 《The Journal of biological chemistry》1988,263(15):7336-7341
2-5A-dependent RNase (RNase L, RNase F) is an enzyme which mediates effects of 2-5A (px(A2'p)nA; x = 2 or 3, n greater than or equal to 2) in cells. 2-5A binding activity present in mouse liver extracts was measured using a 32P-labeled 2-5A derivative. Analysis of Scatchard plots was consistent with a single noninteracting 2-5A binding site with a Ka of 2.5 X 10(10) M-1. Similarly, affinity labeling of proteins with a 32P-labeled 2-5A derivative revealed a single, high-affinity 2-5A-binding protein of Mr 80,000. This 2-5A-binding protein was the only mouse liver protein specifically and consistently eluted by 2-5A from an affinity resin consisting of core(2-5A) covalently attached to cellulose. The 2-5A-eluted protein could degrade polyuridylic acid but not polycytidylic acid. Furthermore, when the 2-5A-eluted protein was electrophoresed into a polyuridylic acid-containing, nondenaturing gel, a band of degraded polyuridylic acid was demonstrated after incubation with 2-5A. There was no band of degraded polyuridylic acid when the elution was performed either in the absence of oligonucleotide or in the presence of low amounts of a closely related analog of 2-5A, p3I2'pA2'pA. Therefore, the Mr 80,000 2-5A-binding protein and the 2-5A-dependent RNase were almost certainly the same protein. Finally, the Mr 80,000 2-5A-binding protein was purified to homogeneity by electroelution from a polyacrylamide gel. 相似文献
997.
M Silverman 《Stain technology》1986,61(3):135-137
Currently available mounting media require solvent evaporation for hardening. This process is slow and often is complicated by the formation of air spaces under the coverslip. Light-polymerizing plastics are introduced as histologic mounting media that eliminate these problems because they harden quickly when exposed to ultraviolet light and, therefore, do not depend upon solvent evaporation. These light-polymerizing plastics are easy to use, permit rapid and controlled hardening, and eliminate the formation of air spaces under the coverslip. The optical characteristics are satisfactory for light and fluorescence microscopy. 相似文献
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M Silverman 《Biochimica et biophysica acta》1976,457(3-4):303-351