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Adsorption of Messenger RNA of Novikoff Hepatoma, Normal Liver, and Regenerating Liver on Complementary DNA-Cellulose Affinity Matrices¹

Department of Pharmacology, Baylor College of Medicine, Houston, Texas 77030

Complementary DNA (cDNA)-oligodeoxythymidylate-celluloses were prepared from cDNA copies of polysomal messenger RNA (mRNA) of Novikoff hepatoma, normal rat liver, and regenerating rat liver. cDNA synthesis with reverse transcriptase was approximately 46% with respect to input mRNA with oligodeoxythymidylate-cellulose primer. The cDNA's of normal liver, regenerating liver, and Novikoff hepatomas were used as affinity matrices for hybridization of different mRNA species. Under the conditions used, degradation of mRNA was not detected. After normalization for homologous hybridization efficiency, 53 and 65% of the Novikoff hepatoma mRNA bound to normal liver and regenerating liver cDNA's. Under these conditions an average of 82% of mRNA of normal liver bound to regenerating liver cDNA, and 92% of regenerating liver mRNA bound to normal liver cDNA. The bound and unbound mRNA's were analyzed by translation in the wheat germ system; 2-D gel analysis of the proteins synthesized in the wheat germ system indicated that the cDNA affinity columns selectively adsorbed some mRNA species.

¹ Supported by the Cancer Research Center Grant CA-10893, awarded by the National Cancer Institute, Department of Health, Education, and Welfare, the Pauline Sterne Wolff Memorial Foundation; the Davidson Fund; and a generous gift from Mrs. J. F. Hutchins.

Received 4/11/77. Accepted 7/14/77.