Purification of human placenta phenylalanine, valine, methionine, glucine, and serine transfer ribonucleic acids.

Biochemistry
M P Amandaraj, B A Roe

Abstract

By using column chromatography on varied media, the purification of several individual tRNAs from human placenta has been achieved. The crude human placenta tRNA was isolated using phenol extraction at pH 4.5 followed by DEAE-cellulose chromatography (B. Roe (1975), Nucleic Acids Res. 2, 21-42) and initially fractionated on BD-cellulose at neutral pH. Subsequent chromatography of the partially purified tRNA using high-speed, high-pressure liquid chromatography on RPC-5 and Aminex A-28 coupled with chromatography on BD-cellulose at acidic pH and on DEAE-Sephadex A-50 significantly shortened isolation time for milligram quantities of several pure tRNA species. Those tRNAs from human placenta obtained in a purity greater than 1.2 nmol/A260 unit are tRNAPhe, tRNAMet(i), tRNAVal(1a), tRNAVal(1b), and tRNAGly(1), while those obtained at purity of at least 0.8 nmol/A260 unit are tRNASer2 and tRNASer3. In addition, the use of Aminex A-28 as a chromatographic system for the isolation of tRNA is discussed.

Citations

Feb 15, 1980·Experientia·M Wozniak, W Dobryszycka
Sep 1, 1977·Proceedings of the National Academy of Sciences of the United States of America·B A Roe, H Y Tsen
Feb 1, 1979·Nucleic Acids Research·B A RoeE Y Chen
Oct 25, 1979·Nucleic Acids Research·R C GuptaK Randerath
Dec 20, 1979·Journal of Chromatography·C T Wehr, S R Abbott
Nov 16, 1976·Biochemistry·R P SinghalG D Novelli

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