Effects of oligonucleotides on the Walker 256 carcinosarcoma DNA polymerase reaction

Abstract

1. The effect of an extract of normal liver on the tumor DNA polymerase (deoxynucleosidetriphosphate: DNA nucleotidyltransferase, EC 2.7.7.7) reaction was found to be similar to the effect produced by nuclease. 2. DNA partially hydrolysed by deoxyribonuclease served as a better primer for the tumor DNA polymerase reaction than either native or heat-denatured DNA. 3. The maximum priming ability of deoxyribonuclease digest for the polymerase reaction was achieved for pancreatic deoxyribonuclease (EC 3.1.4.5) when 20 % of the DNA had been rendered acid soluble, and for splenic deoxyribonuclease (EC 3.1.4.6) or micrococcal deoxyribonuclease (EC 3.1.4.7) when only 1 % hydrolyzed 4. Under conditions of limiting DNA, the polymerase reaction was stimulated by a pancreatic deoxyribonuclease digest of DNA and inhibited by a micrococcal deoxyribonuclease digest. 5. No incorporation of labeled deoxynucleoside triphosphate into oligonucleotide was observed. 6. No incorporation of labeled oligonucleotide into DNA was observed.