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. 1998 Mar 23;140(6):1453–1461. doi: 10.1083/jcb.140.6.1453

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Targeted disruption of the Cx26 gene and transcriptional analysis. (A) Mouse Cx26 wild-type gene with noncoding exon 1 (Ex1) and exon 2 (Ex2) harboring the complete reading frame. (B) Targeting vector: a 1,065-bp fragment of Ex2 was replaced by the neo cassette driven by the PGK promoter. A herpes simplex virus TK cassette was inserted at the 5′ end. The construct was linearized by digestion with NotI, and sticky ends were filled with α-thio dNTP. (C) Mutated, recombinant Cx26 locus and informative restriction fragments, to be compared with the corresponding fragments, in A. The 12- and 7.3-kb fragments represent wild-type and targeted alleles, respectively. The localization of the 3′-external probe and the position of the primers used to genotype embryo yolk sac DNA are shown. PCR primers were selected to generate a 1,118-bp DNA product indicative of the wild-type allele or a 827-bp product due to the targeted Cx26 allele.