Fractionation of Two Diguanylic Acids with Different Internucleotide Linkages and Its Application for Analyses of the Synthetic Products of Ribonuclease T₁ and Spontaneous Reaction

Abstract

1. Fractionation of 2'-5'GpGp and 3'-5'GpGp can be achieved by chromatography on a DEAE-Sephadex column in the presence of 7M urea. Elution was carried out with a linear gradient of NaCl from 0.14 to 0.28M.
2. Diguanylic acid synthesized from G-cyclic-p by RNase T₁ [EC 3. 1. 4. 8] has a 3'-5' phosphodiester linkage.
3. Spontaneous formation of a 2'-5' phosphodiester linkage occurs in the course of purification of G-cyclic-p during the desalting procedure on DEAF-cellulose in the presence of urea.