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Biomedical subjects

G Robey

Publications and source records attributed to G Robey.

4 recordsLinked to original sources

Measurement of binding of terbium to DNA.

This communication demonstrates further that terbium(III) can be used as a probe for DNA. The stoichiometry of terbium binding to DNA was measured by two new methods. In the first method, calf-thymus DNA was titrated with radioactive terbium-160, which is an isotope of the common terbium-159. The resulting DNA-terbium complex was trapped and measured on millipore filters. In the second method, a peak of UV absorption of terbium was found at 219 nm and was used to measure stocichiometry. By both methods, the stoichiometry of binding was one Tb(III) for each three available phosphate groups in DNA. Finally, a rapid method was developed using terbrium-160 to measure the amount of nucleic acid in a solution.

Animals

Terbium as a solid-state probe for RNA.

This paper continues previous work on the analysis of nucleic acid-terbium complexes in the solid state. The fluorescence excitation and emission spectra of the RNA-terbium(III) complex is reported. The fluorescence excitation and emission spectra of both the RNA-terbium(III) and DNA-terbium(III) complexes as trapped on millipore filters is reported. One hundred percent of the DNA combined with terbium was trapped on millipore filters. Deoxyribonucleic acid was recovered from DNA-terbium(III) complexes trapped on millipore filters using SDS-extraction. Energy transfer was shown to occur from the bases in nucleic acids to the terbium ion, whereas the actual binding of terbium to nucleic acids was due to phosphate groups. The relative fluorescence of homopolyribonucleotide-terbium complexes showed that the guanine moiety was responsible for most of the observed fluorescence. Binding studies showed an equal affinity of radioactive terbium for all the homopolyribonucleotides. The fluorescence of solid-state DNA and RNA terbium complexes was used to measure picomole quantities of DNA or RNA.

Animals

Isolation and characterization of a thymic factor.

A protein was isolated from bovine thymus that was shown to accelerate the appearance of hemolysin to sheep erythrocytes in neonatal mice. The isolation procedure consisted of saline homogenization, ammonium sulfate precipitation, and methanol precipitation, followed by chromatography on diethylaminoethyl-cellulose and two chromatographies on Sephadex G-150. The protein was shown to be homogeneous by polyacrylamide gel electrophoresis and by sedimentation velocity and sedimentation equilibrium analytical ultracentrifugation. The molecular weight was determined to be 79,950. Additional physical and chemical characteristics were also determined.

Amino Acids