Figure 1
From: Generation of mice harbouring a conditional loss-of-function allele of Gata6
Generation of a Gata6 conditional null allele. (A) Schematic showing a map of the Gata6 genomic locus and the targeting vector with exons represented by open boxes. The relative position of Southern blot probes (lines), PCR primers (small arrowheads), loxP (large arrowheads) and FRT (ovals) sites, as well as cassettes encoding neomycin phosphotransferase (neo) and diphtheria toxin (DT), are included. Sizes of relevant EcoRI (e), NdeI (n), and PacI (p) restriction endonuclease fragments are shown in kilobase pairs (kb). (B) Southern blot analysis of genomic DNA isolated from ES cells (lanes 1, 2, 8 and 9) or mouse tails (lanes 3–7 and 10). An example of an ES cell line containing a correctly targeted Gata6loxP(FRTneoFRT)allele is shown in lanes 2 and 8. Mice harbouring the modified Gata6 allele were generated from these ES cells (lanes 3 and 10). Exon 2 of Gata6 was deleted (Gata6del) or the neo cassette alone was deleted, leaving Gata6 exon 2 flanked by loxP elements (Gata6loxP), by breeding Gata6loxP(FRTneoFRT)mice to transgenic mice expressing either Cre (lane 6 and 7) or Flp (lane 4) recombinases, respectively. The size of restriction fragments identified by 5' and 3' probes (Fig. 1A) was deduced from their position relative to standard DNA fragments. (C) The genotypes of mice and embryos were also determined by PCR amplification of genomic DNA. Primers were designed that differentiated between the Gata6+ (gt4F/4R; 159 bp), GataloxP(gt4F/4R; 250 bp) and Gata6del(k/o2F/2R; 568 bp) alleles, as well as neo (Neo A/B; 225 bp), flp (Flp F/R; 750 bp), and cre (Cre 1/2; 300 bp) transgenes.