DNA-mediated transfer of the adenine phosphoribosyltransferase locus into mammalian cells.

TLDR

The study demonstrates that DNA‑mediated gene transfer can convert aprt‑deficient mouse cells into the aprt+ phenotype. Transformation was achieved using unfractionated high‑molecular‑weight genomic DNA from Chinese hamster, human, and mouse cells, as well as restriction‑enzyme‑digested rabbit liver DNA. Transformation frequencies ranged from 1 to 10 colonies per 10^6 cells per 20 µg donor DNA, producing transformants with donor‑derived enzymatic activity that were either phenotypically stable or unstable without selective pressure.

Abstract

In this report, we demonstrate the feasibility of transforming mouse cells deficient in adenine phosphoribosyltransferase (aprt; AMP:pyrophosphate phosphoribosyltransferase, EC 2.4.2.7) to the aprt+ phenotype by means of DNA-mediated gene transfer. Transformation was effected by using unfractionated high molecular weight genomic DNA from Chinese hamster, human, and mouse cells and restriction endonuclease-digested DNA from rabbit liver. The transformation frequency observed was between 1 and 10 colonies per 10(6) cells per 20 microgram of donor DNA. Transformants displayed enzymatic activity that was donor derived as demonstrated by isoelectric focusing of cytoplasmic extracts. These transformants fall into two classes: those that are phenotypically stable when grown in the absence of selective pressure and those that are phenotypically unstable under the same conditions.