Fig. 7

Protein trap-induced mutant phenotype affecting the dhx37 gene. (A–D) Bright field images of dhx37^ws0977Tg/+ sibling (A), homozygous dhx37^{ws0977Tg/ws0977Tg} mutant (B), dhx37 ATG morpholino-injected (C), or full-length dhx37 RNA-injected homozygous mutant embryo (D). (E) The Ds insertion in chr8:4765207-4765215 is linked to the mutant phenotype. Black arrows indicate the position of primers used for genotyping PCRs. (F–H) RT-PCR results from homozygous dhx37^{ws0977Tg/ws0977Tg} embryos show that the bcl2 splice acceptor sequence trapped endogenous upstream splice donor sites in the dhx37 gene, producing multiple fusion transcripts (F, G), leading to the disruption of wild-type transcripts (H). (G) The main fusion products, 1 and 2, are out of frame with respect to the reading sequence. Black arrows in the schematic in (F) indicate the positions of primers used for RT-PCR. No evidence of wild-type transcripts was found, and only mutant fusion transcripts were detected. The fusion transcripts always have a 71-bp linker sequence (SA) originated from bcl2 splice acceptor site left between upstream exons and the mCherry reporter sequence. The major fusion transcripts 1 and 2 are out of frame for mCherry coding sequences (G).