Targeting disruption of the <i>Vsig1</i>.

<p>(A) Structure of the wild-type, targeting vector and recombinant allele are shown together with the relevant restriction sites. A 2.5-kb genomic fragment containing exon 1a was replaced by a <i>pgk-neo</i> selection cassette (NEO). The probe used and predicted length of the <i>Eco</i>RI restriction fragment in Southern blot analysis are shown. TK, thymidine kinase cassette; E, <i>Eco</i>RI; X, <i>Xba</i>I; X*, disrupted <i>Xba</i>I site; Xh, <i>Xho</i>I. (B) Blot with <i>Eco</i>RI-digested genomic DNA of recombinant ESC clones was probed with the 3′ probe shown in panel A. The external probe recognized only a 10.7-kb fragment of recombinant allele in <i>Vsig1^−/Y</i> ESCs and a 12.2-kb fragment of the wild-type allele in <i>Vsig1^+/Y</i> ESCs. (C) PCR assay using microsatellite markers was performed to determine the degree of chimerism in the stomachs of chimeric male mice. The 129- and C57-specific fragments were amplified using DNA of the 129/Sv and C57BL/6J mouse strains, and stomach isolated from different chimeric males (Ch).