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Full-Text Articles in Biology
Probing The Alpha-Sarcin Region Of Escherichia-Coli 23s Rrna With A Cdna Oligomer, Gary A. White, Ted Wood, Walter E. Hill
Probing The Alpha-Sarcin Region Of Escherichia-Coli 23s Rrna With A Cdna Oligomer, Gary A. White, Ted Wood, Walter E. Hill
Biological Sciences Faculty Publications
The cause of 50S ribosomal subunit collapse reportedly triggered by hybridization of a 14-base cDNA probe to the alpha-sarcin region of 23S rRNA was investigated by physical measurement of probe-subunit complexes in varying buffer conditions. The results reported here show that this probe was unable to hybridize to its target site in the intact 50S subunit and the physical characteristics of 50S subunits remained unchanged in its presence. Subunit collapse was induced in buffer containing 20mM Tris-HCl (pH 7.5), 600 mM NH4Cl, 1 mM MgCl2, 1 mM DTT, and 0.1 mM EDTA in the absence of probe. The probe bound …
Crystallization Of Trimeric Recombinant Human Tumor Necrosis Factor (Cachectin), M. J. Eck, B. Beutler, G. Kuo, J. P. Merryweather, S. R. Sprang
Crystallization Of Trimeric Recombinant Human Tumor Necrosis Factor (Cachectin), M. J. Eck, B. Beutler, G. Kuo, J. P. Merryweather, S. R. Sprang
Biological Sciences Faculty Publications
Crystals of tumor necrosis factor (TNF) have been obtained in two forms. Rhombohedral crystals grow in 1.8 to 2.0 M ammonium sulfite, pH 7.8 at 21°C, and tetragonal crystals grow in 2.6 M magnesium sulfate, pH 5.5 at 25°C. Analysis of TNF by isoelectric focusing under native and denaturing conditions indicates that TNF molecules exist as trimers in solution. The rhombohedral cachectin crystals belong to space group R3 and have unit cell constants a = b = c = 47.65 Å and α = β = γ = 88.1°. Density determinations and the space group indicate that the unit cell …
Dna Probe Method For The Detection Of Specific Microorganisms In The Soil Bacterial Community, William Holben, Janet K. Jansson, Barry K. Chelm, James M. Tiedje
Dna Probe Method For The Detection Of Specific Microorganisms In The Soil Bacterial Community, William Holben, Janet K. Jansson, Barry K. Chelm, James M. Tiedje
Biological Sciences Faculty Publications
We developed a protocol which yields purified bacterial DNA from the soil bacterial community. The bacteria were first dispersed and separated from soil particles in the presence of polyvinylpolypyrrolidone, which removes humic acid contaminants by adsorption to this insoluble polymer. The soil bacteria were then collected by centrifugation and lysed by using a comprehensive protocol designed to maximize disruption of the various types of bacteria present. Total bacterial DNA was purified from the cell lysate and remaining soil contaminants by using equilibrium density gradients. The isolated DNA was essentially pure as determined by UV spectral analysis, was at least 48 …
Identification Of Defined Sequences In Domain V Of E.Coli 23s Rrna In The 50s Subunit Accessible For Hybridization With Complementary Oligodeoxyribonucleotides, Richard T. Marconi, Walter E. Hill
Identification Of Defined Sequences In Domain V Of E.Coli 23s Rrna In The 50s Subunit Accessible For Hybridization With Complementary Oligodeoxyribonucleotides, Richard T. Marconi, Walter E. Hill
Biological Sciences Faculty Publications
The accessibility of specific sequences in domain V of E. coli 23s rRNA in the 50S subunit to complementary oligodeoxyribonucleotides (cDNA) has been investigated. The apparent percentage of subunits engaged in complex formation was determined by incubation of radiolabeled cDNA probe with 50S subunits, followed by nitrocellulose membrane filtration of the reaction mixtures and measurement of the bound radiolabeled cDNA probes by liquid scintillation counting of the filters. The site(s) of hybridization were determined by digestion of the RNA in the RNA/DNA heteroduplex by RNase H. The results of this study indicated that single-stranded sequences, 2058-2062, 2448-2454, 2467-2483, and 2497-2505 …