Figure 1.

Targeted disruption of the TCTP gene. (A) The structures of the wild-type, targeting vector, and recombinant alleles are shown together with some relevant restriction sites (E, EcoRI; H, HindIII; K, KpnI; N, NdeI). The 5′ and 3′ probes and the predicted length of EcoRI or NdeI restriction fragments in Southern blot analysis are as indicated. (B) Southern blot analysis of the recombinant ES cell clones harboring the “targeted allele.” Genomic DNA extracted from ES cell clones (lanes 1 and 2, nonrecombinants; lanes 3 and 4, clones 248 and 280) was digested with NdeI and probed with the 5′probe. (C) Same as in B except that the genomic DNA was digested with EcoRI and probed with the 3′ probe. The predicted signals for the wild-type (wt) and targeted allele (mt) are as indicated. (D) Representative genotypic analysis of E9.5 embryos harboring the wt (+) or deleted allele (dl or “−”) of the TCTP gene from a TCTP+/− intercross. Genotyping was performed by PCR using primers P1 and P5 for the wild-type (wt, 450 base pairs) and P1 and P4 for the deleted allele (dl, 250 base pairs). (E) Immunoblotting analysis of representative E9.5 embryos with the indicated genotypes using antibodies specific to TCTP or β-actin.