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Flanking loxP sites were introduced to exon 2 and an FRT-flanked neomycin resistance cassette was inserted into the downstream intron via homologous recombination. Deletion of exon 2 and the neo cassette was carried out by transient expression of cre and flp recombinases in correctly targeted ES cells. Western blot analysis of mouse embyonic fibroblasts cultured from E12.5 homozygous mutant embryos confirmed the null allele. It was also shown that the targeted mutation did not interfere with neighboring gene G6pdx. (J:63055)