Analysis of X-ray-Induced HPRT Mutations in CHO Cells: Insertion and Deletions

摘要

Molecular alterations were examined in the hypoxanthine guaninephosphoribosyltransferase (hprt) gene of 41 independent X-ray-induced thioguanine-resistant (TGR) Chinese hamster ovary (CHO) cell clones. Rapid screening of the clones by multiplex polymerase chain reaction (PCR) for the presence or absence of exons revealed that the bases for the mutant phenotype were total gene deletion (26/41), partial gene deletion (4/41), and an insertion (1/41). Southern blot analysis confirmed the deletion data and revealed an additional class of mutants that had a gene disruption but retained all hprt exons (2/41). In addition, the DNA sequence of the insertion mutation was determined.