4 Hogan D.F. et al. (2015) J Vet Cardiol 17(Suppl 1), S306– 17. 5 Li H. et al. (2009) Bioinformatics 25, 1754–60. 6 Li R.H. et al. (2016) J Vet Intern Med 30, 1619–29. 7 Rimmer A. et al. (2014) Nat Genet 46, 912–8. 8 Sherry S.T. et al. (2001) Nucleic Acids Res 29, 308–11. 9 Ziegler S. et al. (2005) Stroke 36, 1394–9. Correspondence: J.A. Stern (jstern@ucdavis.edu) Supporting information Additional supporting information may be found online in the supporting information tab for this article: Table S1 Forward and reverse primer sequences used for PCR amplifications of the target regions in the P2RY1, P2RY12, and CYP2C19 genes. Table S2 Protein alignments of P2Y1, P2Y12, CYP2C19 in cats and other mammalian species around the cat vari- ants p.A236G in P2Y1, p.V34I in P2Y12, and p.P479L in CYP2C19, indicated in bold. doi: 10.1111/age.12668 A nucleotide substitution in the ovine KAP20-2 gene leads to a premature stop codon that affects wool fibre curvature Lingrong Bai* † , Hua Gong †‡ , Huitong Zhou †‡ , Jingzhong Tao* † and Jon G. H. Hickford †‡ *Agricultural College, Ningxia University, Yinchuan 750021, China; † International Wool Research Institute, Faculty of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China; ‡ Gene-Marker Laboratory, Department of Agricultural Sciences, Lincoln University, Lincoln 7647, New Zealand Accepted for publication 20 March 2018 The keratin-associated proteins (KAPs) form a matrix that cross-links the main component of the wool fibre, the ker- atin intermediate filaments. They are therefore believed to play an important role in defining the physical and mechanical properties of the fibre. Although over 80 func- tional KAP genes (KRTAPs) have been identified in humans, only 29 have been found in sheep, 1–3 All of these are polymorphic, and variation in some of these ovine KRTAPs has been associated with wool traits. 1–6 Variation in the gene encoding the high glycine-tyro- sine KAP20-2 protein (KRTAP20-2) has been reported to affect cashmere fibre weight traits. 7 A BLAST search in the Ovine Genome Assembly Oar_v4.0 with the caprine KRTAP20-2 coding sequence (MF973462) identified on chromosome 1 an orthologous open reading frame (NC_019458.2:12334029_123134217), which has seven nucleotide substitutions relative to the caprine sequence. The sequence was located in a region that contains 13 other known KRTAPs (Fig. S1). A comparison of the sheep KRTAP20-2 with the human and caprine homo- logues and with other ovine HGT-KAP genes is shown in Fig. S2. Analysis of a 273-bp PCR fragment containing this open reading frame by single-stranded conformational polymor- phism (SSCP) revealed two nucleotide sequence variants (Fig. S3), corresponding to a single nucleotide polymor- phism (SNP) c.160A>T. This is the only difference between these sequences and the sheep v4.0 sequence, with the A nucleotide being found in that genome assembly sequence. Both sequences have been submitted to GenBank with accession nos. MH071391 and MH071392, for the A- and T-containing sequences respectively. This introduces a pre- mature stop codon that removes the nine C-terminal amino acid residues. In spite of a high frequency of SNPs and inser- tion/deletions in KRTAPs, 1 a nonsense mutation has not been reported to date. The PCR-SSCP analysis of 250 Merino 9 Southdown- cross sheep (from three sire lines) for which wool trait data were available, revealed that the A and T variants occur at a frequency of 71.8% and 28.2% respectively. Analyses using general linear models (GLMs) in MINITAB version 16 indicate that the wool fibre from homozygous AA sheep have a higher mean fibre curvature (MFC) than does the wool fibre from sheep carrying the TT genotype (Table 1). Theoretically, this effect may also reflect linkage between KRTAP20-2 and other genes in the same Table 1 The effect of KRTAP20-2 genotype on various wool traits. Trait Mean Æ SE 1 P AA (n = 143) AT (n = 73) TT (n = 34) GFW (kg) 2.59 Æ 0.04 2.62 Æ 0.06 2.65 Æ 0.07 0.765 CFW (kg) 1.92 Æ 0.03 1.96 Æ 0.04 2.03 Æ 0.06 0.250 Yield (%) 73.9 Æ 0.53 74.9 Æ 0.82 76.2 Æ 1.09 0.149 MSL (mm) 86.9 Æ 1.04 88.1 Æ 1.59 87.7 Æ 2.12 0.797 MSS (N/ktex) 24.5 Æ 0.66 22.8 Æ 1.02 25.8 Æ 1.35 0.797 MFD (lm) 19.2 Æ 0.15 18.7 Æ 0.24 18.9 Æ 0.31 0.091 FDSD (lm) 3.97 Æ 0.05 3.87 Æ 0.08 3.83 Æ 0.11 0.387 CVFD (%) 20.5 Æ 0.19 20.7 Æ 0.29 20.1 Æ 0.39 0.459 MFC ( o /mm) 84.5 Æ 1.19 A 80.5 Æ 1.82 AB 74.1 Æ 2.42 B <0.001 PF (%) 2.42 Æ 0.28 1.80 Æ 0.43 1.93 Æ 0.57 0.404 GFW, greasy fleece weight; CFW, clean fleece weight; MSL, mean staple length; MSS, mean staple strength; MFD, mean fibre diameter; FDSD, fibre diameter standard deviation; CVFD, coefficient of varia- tion of fibre diameter; MFC, mean fibre curvature; PF, prickle factor (percentage of fibres over 30 lm). 1 Estimated marginal means, standard errors of these means and P- values derived from GLMs. Means within rows that do not share a superscript uppercase letter (e.g. A and B) were different at P < 0.01. 357 © 2018 Stichting International Foundation for Animal Genetics, 49, 349–358 Brief Notes