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J. Biochem, 1977, Vol. 81, No. 5 1247-1252
© 1977 Japanese Biochemical Society

research-article

Separation of Newly-Synthesized RNA by Organomercurial Agarose Affinity Chromatography1

Masao ONO and Masaya KAWAKAMI

Molecular Biology Laboratory, School of Medicine, Kitasato University Sagamihara, Kanagawa 228

Mouse myeloma cells, MOPC-31C, were incubated in the presence of 2-thiouridine and newly synthesized RNA which appeared to contain 2-thioUMP as a constituent was separated from preexisting RNA by affinity chromatography using organomercurial agarose as a support. Both pH and salt concentration greatly affected the specific adsorption of the newly-synthesized RNA on the column. Under optimal conditions the rate of adsorption of the newly-synthesized RNA on the column was proportional to the logarithmic concentration of 2-thiouridine in the culture medium. Furthermore, at a given concentration of 2-thiouridine in the medium, a shorter incubation period caused a reduction of the rate of RNA adsorption on the column. The molecular size distributions of both total RNA and the adsorbed fraction, synthesized during 30 mm in the presence of 2-thiouridine, were similar to that of RNA synthesized in the absence of the drug.

1This study was supported in part by a grant from the Ministry of Education, Science and Culture of Japan.


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