Fig. 1

Gene targeting of the mouse protrudin locus. a Schematic representation of the domain structure for the protrudin open reading frame (ORF), the genomic structure of the protrudin gene locus, the targeting vector, the wild-type (WT) allele, and the mutant (KO) allele after homologous recombination. A genomic fragment including exons I and II of the protrudin coding region (red) was replaced with a phosphoglycerate kinase (PGK)–neo cassette. UTR untranslated region, RBD Rab11 binding domain, TM transmembrane domains, HP hairpin domain, FFAT two phenylalanines in an acidic tract, CC coiled-coil domain, FYVE Fab1-YOTB-Vac1-EEA1 domain, tk thymidine kinase gene, neo neomycin resistance gene, E1 EcoR1 site. b Southern blot analysis of EcoRI-digested genomic DNA from mice of the indicated protrudin genotypes (PRT) with the probes shown in (a). The 10- and 6.5-kbp bands corresponding to the WT and KO alleles, respectively, are indicated. c PCR analysis of genomic DNA from mice of the indicated protrudin genotypes. The 440- and 340-bp bands corresponding to the WT and KO alleles, respectively, are indicated. d Quantitative RT-PCR analysis of mRNA derived from exons II or X of the protrudin gene in mouse embryos of the indicated protrudin genotypes. Data are means + SEM (n = 3). ***P < 0.001 (Student’s t test)