Serial transfer of a human gene to rodent cells by sequential chromosome-mediated gene transfer.

Abstract

The human hypoxanthine phosphoribosyltransferase (IMP:pyrophosphate phosphoribosyltransferase, EC 2.4.2.8) gene (hprt) was serially transferred to mouse cells and then to Chinese hamster fibroblasts by 2 cycles of metaphase chromosome isolation and incubation with recipient cells. Human metaphase chromosomes were incubated with mouse A9 cells deficient in hypoxanthine phosphoribosyltransferase, and independent colonies expressing the human species form of this gene were isolated in a selective medium. Metaphase chromosomes isolated from 2 of these clonal lines were incubated with Chinese hamster fibroblasts deficient in hypoxanthine phosphoribosyltransferase; 5 resulting independent colonies again expressed the human species of this gene. The transfer frequencies in the 2 cycles of chromosome-mediated gene transfer were similar (about 10-7). The transferred human chromosome fragment is closely associated with the chromosomes of the mouse A9 cells and it is probably integrated into the chromosomal DNA of the recipient cell.