Identification of the ovine KAP11-1 gene (KRTAP11-1) and genetic variation in its coding sequence
Identification of the ovine KAP11-1 gene (KRTAP11-1) and genetic variation in its coding sequence
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DOI：
10.1007/s11033-011-0697-2
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发表时间：
2011-11-01
影响因子：
2.8
通讯作者：
Hickford, Jon G. H.
中科院分区：
文献类型：
--
作者：
Gong, Hua;Zhou, Huitong;Hickford, Jon G. H.
角蛋白相关蛋白（KAP）是羊毛纤维的结构组分，并形成角蛋白中间丝（KIF）之间的基质。编码高硫蛋白KAP 11 -1的基因已在人类、牛和小鼠中被鉴定，但尚未在绵羊中被鉴定，尽管羊毛具有经济重要性。在这项研究中，使用基于牛KAP 11 -1基因序列的引物进行PCR，用绵羊DNA产生预期大小的扩增子。应用PCR-SSCP技术对260只绵羊进行分析，共检测到6种不同的PCR-SSCP带型。每只绵羊都有一种或两种带型的组合，表明它们对该基因是纯合的或杂合的。扩增子的测序证实了六个DNA序列的存在。所有这些都是独特的，并且与来自牛、人和小鼠的KRTAP 11 -1序列的同源性最大，表明它们来源于绵羊KAP 11 -1基因并且是等位变异体。绵羊KAP 11 -1基因开放阅读框为477个核苷酸，编码159个氨基酸。推测的蛋白质富含丝氨酸、半胱氨酸和苏氨酸，分别占18.2- 18.9mol%、12.6mol%和12.0mol%。其中，大约20个丝氨酸和苏氨酸残基可能被磷酸化。鉴定了5个核苷酸取代，1个是非同义的，将导致潜在磷酸化位点的氨基酸变化。在KRTAP 11 -1中发现的遗传变异可能影响其表达、蛋白质结构和/或翻译后修饰，从而影响羊毛纤维结构和羊毛性状。
Keratin-associated proteins (KAPs) are a structural component of the wool fibre and form the matrix between the keratin intermediate filaments (KIFs). The gene encoding high sulphur-protein KAP11-1 has been identified in human, cattle and mouse, but not yet in sheep, despite the economic importance of wool. In this study, PCR using primers based on the cattle KAP11-1 gene sequence produced an amplicon of the expected size with sheep DNA. Upon using PCR-Single Stranded Conformational Polymorphism (PCR-SSCP) analysis in 260 sheep, six different PCR-SSCP patterns were detected. Either one or a combination of two banding patterns was observed for each sheep, suggesting they were either homozygous or heterozygous for this gene. Sequencing of the amplicons confirmed the occurrence of six DNA sequences. All of these were unique, and the greatest homology was with KRTAP11-1 sequences from cattle, human and mouse, suggesting that they were derived from the ovine KAP11-1 gene and were allelic variants. The ovine KAP11-1 gene had an open reading frame of 477 nucleotides encoding 159 amino acids. The putative protein was rich in serine, cysteine, and threonine which account for 18.2-18.9, 12.6 and 12.0 mol%, respectively. Of these, approximately 20 of the serine and threonine residues might be phosphorylated. Five nucleotide substitutions were identified, and one was non-synonymous and would result in an amino acid change at a potential phosphorylation site. The genetic variation found in KRTAP11-1 may influence its expression, protein structure, and/or post-translational modifications, and consequently affect wool fibre structure and wool traits.