Functional hemizygosity for the MDH2 locus in Chinese hamster ovary cells

Isolation of electrophoretic mobility shift mutants for a large number of enzyme loci in CHO cells has allowed the identification of many genes which are functionally hemizygous. To gain further insight into the nature of hemizygosity in CHO cells and the mechanisms by which it has arisen, we are attempting to determine whether hemizygous gene loci are clustered in a few localized chromosomal regions in CHO or are more generally distributed throughout the genome. Isozyme analysis of a series of CHO electrophoretic mobility shift mutants for MDH2 (malate dehydrogenase 2, EC 1.1.1.37) revealed that this locus is functionally hemizygous in CHO cells, but the locus could not be mapped by conventional approaches because of the similar electrophoretic mobilities of Chinese hamster and mouse MDH2 isozymes. Construction of intraspecific CHO X CHO hybrids using electrophoretic mobility shift mutants with secondary, selectable drug-resistance markers allowed us to determine that MDH2 is not closely linked to any previously mapped hemizygous marker loci in CHO, but is linked to alleles for two dizygous gene loci, PGM3 and APRT, on CHO chromosome Z7. A possible genetic basis for hemizygosity of the MDH2 locus in CHO cells is discussed.