High-frequency deletion event at aprt locus of CHO cells: detection and characterization of endpoints.

Two mechanisms are implicated in generating recessive drug resistance mutants at the adenine phosphoribosyltransferase (aprt) locus of Chinese hamster ovary (CHO) cells, one of which is a spontaneous high-frequency deletion of the entire gene. We have isolated and mapped a 19-kb fragment carrying aprt and its flanking sequences. A Southern blot study of 198 independent deletion mutants revealed that two different mutants have one of their breakpoints within the 19-kb region analyzed. One of these has an upstream breakpoint which could be narrowed down to a 4-kb fragment containing repetitive sequences. The other mutant has a breakpoint within a 410-bp sequence located 8.5 kb downstream of the aprt gene and which carries several elements similar to those signaling V-(D)-J joining in immunoglobulin and T-cell receptor gene rearrangements. In each case the other breakpoint lay outside of the analyzed region. These results support the previous indications that the deletions created by this spontaneous event are large.