Figure 3.
Figure 3. The qmc551 and sa11633 alleles of gfi1aa. (A) Genomic maps of 2 wild-type and 2 mutant alleles of gfi1aa, including information about transcripts and their splice products. The 2 wt alleles differ in intron 1. Intron 1 of allele wt1 carries a hAT-Tol2 family transposon called DNA8-9_DR whose target site was duplicated upon integration. The duplicated sequences flank the transposon as direct repeats. In the qmc551 allele, the gene trap transposon was inserted into the DNA8-9_DR transposon of the wt1 allele. Please note that most gfi1aaqmc551Gt transcripts terminate behind the gfp reading frame. Splicing between the splice donor at the end of exon 1 and the splice acceptor on the gene trap transposon allows GFP expression from the qmc551 allele.37 The wt2 and sa11633 alleles do not possess transposon insertions in intron 1. Their intron 1 sequence is considerably shorter. The sa11633 allele carries a nonsense mutation in exon 4. Detailed annotations are provided in the inset. (B) The diagram shows the full-length wt Gfi1aa protein with all its functional domains and the truncated Gfi1aa protein encoded by the sa11633 allele. (C) PCR of exon 4 sequences and subsequent DNA sequencing are required to distinguish wt and sa11633 alleles of gfi1aa. The panels show Sanger DNA sequencing reads of the template strand of the wt and sa11633 alleles of gfi1aa. The coding sequence has been added below the sequence read. It shows that the mutation converts the serine145 codon to a premature stop codon. (D) PCR-based genotyping of wt and qmc551 alleles of gfi1aa. In lanes 2 to 7, PCR products are shown that were amplified on genomic DNA samples that did (+) or did not (−) contain the respective gfi1aa allele. The expected wt1, wt2, and qmc551 fragments are 1322, 477, and 491 bp in size. Information on primer sequences is provided in supplemental Table 8. (E-F) Lateral views of embryos stained by WISH. Yellow and green arrows point at gene expression in inner ear hair cells and in the ventral wall of the dorsal aorta, respectively. Numbers of embryos analyzed are provided on the panels.

The qmc551 and sa11633 alleles of gfi1aa. (A) Genomic maps of 2 wild-type and 2 mutant alleles of gfi1aa, including information about transcripts and their splice products. The 2 wt alleles differ in intron 1. Intron 1 of allele wt1 carries a hAT-Tol2 family transposon called DNA8-9_DR whose target site was duplicated upon integration. The duplicated sequences flank the transposon as direct repeats. In the qmc551 allele, the gene trap transposon was inserted into the DNA8-9_DR transposon of the wt1 allele. Please note that most gfi1aaqmc551Gt transcripts terminate behind the gfp reading frame. Splicing between the splice donor at the end of exon 1 and the splice acceptor on the gene trap transposon allows GFP expression from the qmc551 allele.37  The wt2 and sa11633 alleles do not possess transposon insertions in intron 1. Their intron 1 sequence is considerably shorter. The sa11633 allele carries a nonsense mutation in exon 4. Detailed annotations are provided in the inset. (B) The diagram shows the full-length wt Gfi1aa protein with all its functional domains and the truncated Gfi1aa protein encoded by the sa11633 allele. (C) PCR of exon 4 sequences and subsequent DNA sequencing are required to distinguish wt and sa11633 alleles of gfi1aa. The panels show Sanger DNA sequencing reads of the template strand of the wt and sa11633 alleles of gfi1aa. The coding sequence has been added below the sequence read. It shows that the mutation converts the serine145 codon to a premature stop codon. (D) PCR-based genotyping of wt and qmc551 alleles of gfi1aa. In lanes 2 to 7, PCR products are shown that were amplified on genomic DNA samples that did (+) or did not (−) contain the respective gfi1aa allele. The expected wt1, wt2, and qmc551 fragments are 1322, 477, and 491 bp in size. Information on primer sequences is provided in supplemental Table 8. (E-F) Lateral views of embryos stained by WISH. Yellow and green arrows point at gene expression in inner ear hair cells and in the ventral wall of the dorsal aorta, respectively. Numbers of embryos analyzed are provided on the panels.

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