Induced segregation of human syntenic genes by 5-bromodeoxyuridine + near-visible light

Abstract

Chromosome-breaking agents have been used in two different procedures for promoting segregation of syntenic genes on human chromosome 12. In method A, a human-Chinese hamster cell hybrid containing the single human chromosome 12 was treated either with 5-bromodeoxyuridine BrdU + near-visible light or with X-rays. In method B, normal human fibroblasts were treated with BrdU + near-visible light followed by their fusion with a Chinese hamster glycine-requiring cell mutant CHO- K1/gly⁻A. Since the human complementing gene for serine hydroxymethyltransferase, an enzyme deficient in gly⁻A, lies on human chromosome 12, only those hybrids retaining that chromosome can survive the glycine-free medium. Clones isolated from both procedures were analyzed for the loss or retention of four other syntenic genes on chromosome 12, TPI, GAPD, LDH_B, and PepB. The results demonstrate that method B is much more effective in generating clones with extensive marker losses. In addition, the segregation pattern and frequency obtained in this study provided information on the linear order of TPI and GAPD on chromosome 12.