Figure 1: Targeted disruption of the Nczf gene.

(A) The Nczf locus containing 6 exons and a portion of the 5′ and 3′ flanking regions is shown at the top. The targeting vector was designed to replace a DNA segment through exon 4 to exon 6 by a neomycin resistant gene cassette (NEO). A diphtheria toxin A gene cassette (DT-A) was used for a negative selection. The recombinant allele, resulting from homologous recombination, is depicted in the bottom panel. The coding exons or portions of exons are depicted by filled boxes and the open boxes denote the non-coding portions. The positions of the primers (P1 and P2) used for genotyping by PCR analysis are also indicated. The orientation of both NEO and DT-A was the same as that of Nczf. (B) PCR analysis of genomic DNA extracted from E8.5 mice. A primer set (P1 and P2) was used to detect the wild type (WT) allele (400 bp) and the primer set (P3 and P4) was used to detect Nczf knockout (KO) allele (800 bp). Whole image of the gel electrophoresis data is shown in supplementary information.