Figure 1

Rbm19^{GtXC768/GtXC768}gene-trap characterization. (A) The insertion site of the gene trap vector is in intron 16 (*), resulting in loss of RRMs 5 and 6, along with the addition of a 116 kD β-geo protein. The exact insertion site is shown by the sequences flanking the gene trap insertion. The nucleotide number corresponds to the sequence available at NCBI accession NT_078458.6. The bold nucleotides represent the primer sequences we used to detect the wild-type Rbm19 allele. (B) RT-PCR shows no detectable Rbm19⁺ transcript in the mutant E3.5 embryos using Rbm19-specific primers made to exons flanking the insertion site (red arrows, panel A). The expected product was detected when using an Rbm19-specific forward primer and β-geo reverse primer in the mutant embryos. (C) Western blot for RBM19 (amino-terminal epitope [14]) shows the expected 191 kD fusion protein band in the heterozygous mouse intestine (arrow), but not in homozygous wild-type. (D) ³²P-labeled PCR genotyping of E3.5 embryos from Rbm19^GtXC768/+ intercrosses. The embryos were genotyped by a duplex PCR strategy. Rbm19-specific primers amplify a 296-bp DNA fragment across the insertion site (panel A, WT), which is disrupted by the genetrap vector in the mutant. β-geo-specific primers amplify a 507-bp fragment from the gene trap vector (panel B, Rbm19^GtXC768.)