Generation of TMPRSS11A-deficient mice.

<p>A. Schematic structure of the gene targeting replacement vector (top), wildtype <i>Tmprss11a</i> gene (middle), and targeted <i>Tmprss11a</i> gene (bottom). Position of Eco RI restriction enzyme cleavage sites used for digestion of genomic DNA for Southern blot analysis, position of Southern blot probe (bar), and positions of primers used for analysis of wildtype and mutant <i>Tmprss11a</i> alleles (arrows) and transcripts (arrowheads) are indicated. B. Southern blot hybridization of Eco-RI digested DNA from a control (lane 1) and a targeted (lane 2) embryonic stem cell clone. The positions of wildtype (14.8 kb) and mutant <i>Tmprss11a</i> (6.5 kb) alleles are indicated on the right. Positions of molecular weight markers (kb) are indicated at left. C. PCR analysis of tail biopsy DNA of offspring from interbred <i>Tmprss11a^+/−</i> mice. The position of wildtype (496 bp) and mutant <i>Tmprss11a</i> (266 bp) alleles are indicated. D. RT-PCR analysis of <i>Tmprss11a</i> mRNA transcripts from tongue of <i>Tmprss11a^+/+</i> (lane 1) and <i>Tmprss11a^−/−</i> (lane 2) mice using the exon 7-flanking primer pair indicated with arrowheads in A. Lanes 3 and 4, no reverse transcriptase and no RNA added to the reactions, respectively. Bottom panel. Amplification of <i>Gapdh</i> mRNA demonstrating the integrity of the cDNA preparation. E. RT-PCR amplication of <i>Tmprss11a</i> mRNA transcripts from tongue of <i>Tmprss11a^+/+</i> (lanes 1, 6, 11, and 16) and <i>Tmprss11a^−/−</i> (lane 2, 7, 12, and 17) mice using primer pairs capable of amplifying exons 2–9 (lanes 1–5), 2–5 (lanes 6–10), 8–9 (lanes 11–15), and <i>Gapdh</i> (lanes 16–20). Reverse transcriptase was omitted from the reactions in lanes 3, 4, 8, 9, 13, 14, 18, and 19. No RNA was added to reactions in lanes 5, 10, 15 and 20. Transcript size is indicated left.