PMC:1853120 / 44193-46175 JSONTXT

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In the resulting vector, the second and third exons of Dmrt7 are flanked by loxP sites (floxed). The Dmrt7-containing portions of pDZ169 were completely sequenced.\npDZ169 was linearized with PmeI and electroporated into CJ7 ES cells (originally derived from the 129S1 strain). Three homologous recombinants were identified from 296 G418-resistant colonies by Southern hybridization by use of a DNA probe from the sequences upstream of exon 1 to screen genomic DNA digested with EcoRI. Homologous recombination was confirmed on both ends of the targeted region by Southern hybridization. Probes for Southern hybridization were made by PCR using primers DM5S10/DM5S11 (5′ probe) and DM5PR1/DM5PR2 (3′ probe), listed below. Two targeted ES cell clones containing the floxed allele Dmrt7neo were injected into C57Bl/6 blastocysts to generate chimeras. Chimeric males were bred with C57Bl/6 females to generate heterozygotes carrying Dmrt7neo. Dmrt7+/Dmrt7neo females were bred with male β-actin-Cre transgenic mice to delete the floxed sequences and generate heterozygous Dmrt7− /+ deletion mutants, which were interbred to generate homozygous Dmrt7−/− 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