Incorporation of deoxynucleotides into DNA by diethylaminoethyldextran-treated lymphocytes
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Abstract:
In the presence of diethylaminoethyldextran cultured human lymphocytes will utilize deoxynucleotides for the synthesis of DNA, whereas in the absence of diethylaminoethyldextran no incorporation of deoxynucleotides is detected. Labeled deoxynucleoside mono-, di-, and triphosphates are incorporated into DNA at approximately the same rate. Deoxynucleotide incorporation is essentially linear for 10 min but continues at a gradually diminishing rate for an additional 20 to 50 min. The initial rate of DNA synthesis is at least 20 to 40% of the in vivo rate, and in those cells that are in S phase 0.7-1.5% of the DNA is synthesized. By the three properties examined (restriction to S phase, semiconservative mode, and initial product in short chains), DNA synthesis in diethylaminoethyldextran-treated cells resembles DNA synthesis in vivo.