Additional file 6.
a. Alignment of tammar Ptch2 protein sequence with four eutherian mammals. Dark shading indicates agreement in at least 60% of the sequences, light shading indicates amino acid similarity to consensus. 70 amino acid stretch maintained in Tammar is italicized. Double dashed areas represent putative trans-membrane binding domains, with species showing highest sequence identity indicated in parentheses. Any conserved domains are mentioned above the relative sequence. b. Alternative splice variants of PTCH2. Primers were designed to span the region corresponding to exons 18-22 of the human PTCH2 gene. RT-PCR was carried out in day 3, 7 and 14 post partum testes. Day 3 PCR produced four bands of ~1.2 Kb, 1.05 Kb, 950 bp and 860 bp. We sequence verified that the 1.2 Kb fragment was the full-length transcript and that the 950 bp transcript was a Δ-21a PTCH2 isoform. The identity of the missing exons in the 1.05 Kb and 860 bp fragments is shown in Additional File 9. These slice variants were developmentally regulated, with the smaller two isoforms not seen in the day 7 or 14 testis and the larger two isoforms appear to change in their relative abundance between stages.
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O'Hara et al. BMC Developmental Biology 2011 11:72 doi:10.1186/1471-213X-11-72