FIG. 1.

Generation and tissue-specific expression of the Wt1-Dmrt1 transgene. A) Generation of the Wt1-Dmrt1 transgene. The rat Dmrt1 cDNA was inserted in frame with the translational start ATG of mouse Wilms tumor gene 1 (Wt1) within the yeast artificial chromosome YAC620mWt1. Top: YAC620mWt1; exons = boxed with numbers, URA/Acentric, and TRP/Centric = the pYAC4 acentric vector arms with uracil and tryptophan selectable markers, respectively. Middle: targeting vector, pBKS-Wt1-Dmrt1-HPRT1-LTS2. Includes the rat Dmrt1 cDNA, exons 8 and 9, intron 8 and polyadenylation site of HPRT1, lysine 2 selectable marker (LYS2), 825 bp of Wt1 sequence 5′ of translational start site (Wt15′), and 776 bp of Wt1 sequence 3′ of translational start site (Wt13′). Bottom: Wt1-Dmrt1 transgene after homologous recombination in yeast (AB1380). B and C) RT-PCR of RNA prepared from different organs from wild-type (wt superscript) and transgenic (tg superscript) mice. B) Transgene expression in the kidneys (K) and ovaries (O) and testis (T) of newborn littermates. M and F subscripts indicate male and female, respectively. C) Transgene expression in P15 male mice. Tissues are denoted as testis (T), brain (B), heart (H), spleen (Sp), stomach (St), lung (Lu), kidney (K), and liver (Li). The plasmid pBKS-Wt1 5′-Dmrt1-HPRT1-LYS2-Wt1 3′ (P) was included as positive control and water (H₂O) as negative control. cDNA was synthesized in the presence (+) or absence (−) of reverse transcriptase. A ∼450-bp band denotes cDNA amplification, while the ∼1105-bp band bottom is amplified genomic DNA.