Chromosome-mediated gene transfer of HPRT and APRT in an intraspecific human cell system.
Chromosome-mediated transfer of genes between human cell lines was accomplished using HeLa cells as chromosome donors and HT1080 fibrosarcoma lines as recipients. This report describes the intraspecific transfer of two genetic markers, hypoxanthine-guanine-phosphoribosyltransferase (HPRT+) and adenine phosphoribosyltransferase (APRT+). The isolation and characterization of the necessary enzyme-deficient (HPRT- and APRT-) recipient HT1080 cell lines are also described. The chromosome-mediated gene transfer was carried out using a modification of the procedure of Miller and Ruddle, including treatment of the donor chromosomes with calcium phosphate and subsequent exposure of the recipient cells of dimethyl sulfoxide. In experiments to optimize this procedure for HT1080 cell recipients, we found that a brief (2-min) exposure to high DMSO concentration (20%) was effective for enhancing transfer efficiencies in this system. Transfer frequencies (transferents per recipient cells assayed) averaged approximately 1 x 10(-6) for HPRT+ and were greater than 2 x 10(-6) for APRT+.
D000228:Adenine Phosphoribosyltransferase / Q000235:genetics*; D002460:Cell Line; D002877:Chromosomes, Human / Q000502:physiology*; D005354:Fibrosarcoma; D005796:Genes; D006367:HeLa Cells / Q000201:enzymology; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics*; D009154:Mutation; D010430:Pentosyltransferases / Q000235:genetics*
D010430:Pentosyltransferases; D000228:Adenine Phosphoribosyltransferase; D007041:Hypoxanthine Phosphoribosyltransferase