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Nucleic Acids Research, 1980, Vol. 8, No. 14 3073-3086
© 1980

ENZYMOLOGY

2'5' Obgoadenylate synthetase, an interferon induced enzyme: direct assay methods for the products, 2'5' ohgoadenylates and 2'5' co-oligonucleotides

J. Justesen, D. Ferbus and M.N. Thang

Institut de Biologie Physico-Chimique 13, rue Pierre et Marie Curie, 75005 Paris, France

Received June 3, 1980. The interferon induced enzyme 2'5' oligoadenylate synthetase produces 2'5' pppA(pA)n the first discovered natural nucleotide with a 2'5' linkage. We describe a direct assay of this enzyme based on separation by thin layer chromatography (TLC) of the substrate ATP and the products 2'5'pppA(pA)n (n ≥ 1). This technique presents obvious advantages compared to the currently used methods. Moreover the enzyme uses other nucleotides as substrates forming co-oligonucleotides 2'5'pppA(pA)n pN (N= U,G,C,dA,dG,dT and dC). Additional procedures are described using different developing solvent systems for the separation of the core-2'5' oligonucleotides (2'5 A(pA)npN) containing AMP-residues entirely and those with another nucleotide at the 2'end.


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