Figure 1 (A) Representation of the position of the
sv15 and
im9 mutations in
tsp-15, and the effects of the mutations on the splicing of intron 4. The
sv15 mutation alters the splice donor site, and a cryptic splice donor in the middle of intron 4 is often used (Moribe et al. 2004). The intragenic
im9 mutation abolishes the cryptic donor site, resulting in reuse of the original donor site and partial recovery of the wild-type transcript. (B) Design of qRT-PCR analysis using TaqMan probe for detecting correctly spliced
tsp-15 transcript. The positions of primers and TaqMan MGB probe are indicated. R, reporter; NFQ, nonfluorescent quencher; MGB, minor groove binder. (C) Relative expression levels of correct
tsp-15 transcripts. Data are mean RQ value and error bars indicate RQmin/RQmax (confidence set at 95%).
ok881 is a null allele of
tsp-15 having a deletion in the
tsp-15 coding region (Moribe et al. 2012). (D) Representation of splice pattern of
tsp-15 cDNA from
sv15im9. Among 12
tsp-15 cDNA subclones, we found 3 correctly spliced products, 3 containing read-through of intron 4, and 6 aberrant transcripts using -GT- in exon 4 as a cryptic splice donor.