GENETICS OF HUMAN CELL LINES, IV. DNA-MEDIATED HERITABLE TRANSFORMATION OF A BIOCHEMICAL TRAIT

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Abstract
The treatment of the IMP-pyrophosphorylase (IMPPase)-deficient human cell line, D98/AH-2, with DNA isolated from IMPPase-positive D98S or D98/AG cells resulted in the appearance of IMPPase-positive genetically transformed cells, which were detectable under highly selec -tive conditions. DNAse but not RNAse treatment abolished the transforming activity. The yield of transformants was a linear function of the transforming DNA concentration up to the level of about 20 [mu]g DNA/ml, reaching a plateau of approximately 4 X 10-4 transformations per recipient cell at 150 [mu]g DNA/ml. Transforming activity banded in the CsCl equilibrium gradient more sharply than the total DNA mass. DNA isolated from the D98/AH-2 recipient cells or from the cells of other mammals did not transform per se, but interfered with the active transformation process. No recombinations between intact cells were detected, using aminopterin and 8-azahypoxanthine resistance as selective markers.