Identification of the ovine keratin-associated protein KAP1-2 gene (KRTAP1-2).

Keratin-associated proteins (KAPs) are a major component of wool and other keratin-containing tissues. While four KAP1-n proteins have been identified in sheep, only three genes have been described encoding KAP1-1, KAP1-3 and KAP1-4. Here, we used a sequence conserved across the known KAP1-n genes to search the inaugural Ovine Genome Sequence (v1.0) and identified a new KAP1-n sequence on chromosome 11. PCR amplification of this sequence revealed an open reading frame of 474-bp that putatively encodes a polypeptide sequence very similar to the previously described ovine KAP1-2 protein and suggests that the newly identified sequence represents the previously unidentified KAP1-2 gene (KRTAP1-2). Its expression in skin was confirmed by PCR, and the mRNA was localized to the cortex of the mid-keratinization zone of a growing wool fibre using a gene-specific probe and in situ hybridization. PCR-SSCP analysis of KRTAP1-2 revealed nine unique banding patterns representing nine different DNA sequences. One sequence was identical to, and the other eight were homologous to, the sequence identified above, suggesting that they were allelic variants of ovine KRTAP1-2. There were ten single nucleotide substitutions identified, although only three of these were non-synonymous and would potentially result in amino acid changes. The variation identified here may influence the expression or protein structure of KAP1-2 and consequently wool structure and wool traits.