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Stimulation of Oligodeoxyribonucleotide Synthesis by Cytoplasmic Factors in the Isolated Nuclei of Leukemic Cells¹

Nova University, Life Sciences Center, Fort Lauderdale, Florida 33314

Nuclei were isolated by hypotonic shock and homogenization from freshly drawn, nonstimulated leukemic leukocytes and were pulse labeled with deoxythymidine triphosphate-³H. Labeled oligodeoxyribonucleotides were separated from the bulk of DNA and proteins by membrane cone filtration and were further purified by alkaline hydrolysis and diethylaminoethyl cellulose chromatography. In parallel experiments, cytoplasmic factors, obtained after removal of nonsoluble material from the cytoplasm by high-speed centrifugation, were included. More than a twofold increase of label incorporation into oligodeoxyribonucleotides was observed.

¹ Nova University and, in part, the leo Goodwin Institute for Cancer Research supported this work. The Packard 2425 liquid scintillation counter, used in these experiments, was purchased from a grant given by the American Cancer Society, Florida Division, Inc. (F72N-1), and supplemented by Nova University.

Received 1/29/73. Accepted 6/15/73.