Nucleic Acids Research, 1975, Vol. 2, No. 6 853-864
© 1975
Articles |
An Improved method for the purification of tRNA by chromatography on dlhydroxyboryl substituted cellulose
*Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut 06520 and Department of Biological Sciences, Purdue University Lafayette, Indiana 47907
Received April 4, 1975. An Improved method for the rapid separation of aminoacyl-tRNA from tRNA by chromatography on dihydroxyboryl-substituted cellulose has been developed. The method relies on the selective binding of unacylated tRNA to the cellulose support containing dlhydroxyboryl groups. This binding is the result of complex formation between the cis-diol group of the 3'-terminal ribose in tRNA and the dihydroxyboryl groups immobilized on the resin. Aminoacyl-tRNA cannot undergo borate complex formation and is not retained on the resin. The separation is carried out at near neutral pH values ensuring stability of the aminoacyl ester linkage. The aminoacyl-tRNAs are obtained in very high purity. Aminoacyl-tRNA species containing the modified nucleoside Q are also retained on dihydroxyboryl cellulose. Conditions for Isolating all Q base containing tRNA species from unfractionated tRNA are described.
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