Characterization of gpt deletion mutations in transgenic Chinese hamster cell lines.
The transgenic cell lines G12 and G10, each with a bacterial gpt gene stably integrated at a single but different position in the Chinese hamster genome, were evaluated for deletion of the gpt transgene following exposures to several clastogens. More than 150 independently cloned G12 and G10 6-thioguanine-resistant mutants have been characterized by polymerase chain reaction (PCR) amplification and Southern blots in this study. Despite differences in the integration sites for the gpt gene in the G12 and G10 cells, PCR amplification of the gpt gene from both cell lines can be performed using the same single set of primers. By PCR deletion screening, about 20% of recovered spontaneous 6-thioguanine resistant (6TG) gpt G12 mutants had deleted the transgene, whereas the deletion mutant frequency was increased to about 50% of the X-ray- and bleomycin-induced G12 mutants. In contrast, both spontaneous and induced deletion frequencies are considerably higher for the G10 cell line. Among spontaneous G10 mutants, up to 50% have deleted the gpt transgene, whereas almost all of the X-ray- and bleomycin-induced G10 mutants have lost the integrated gene sequence. These results are discussed in the context of the transgene integration sites and the influences of the surrounding genome that may render certain genetic regions prone to deletion.
Klein
C B
CB
New York University Medical Center, Nelson Institute of Environmental Medicine, Tuxedo 10987, USA.
Su
L
L
Singh
J
J
Snow
E T
ET
D000410:Alanine Transaminase / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D017353:Gene Deletion; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D019076:Transgenes / Q000235:genetics*
D000410:Alanine Transaminase