麦洼牦牛毛色候选基因<em>STX17</em>的克隆及组织表达分析

doi:10.3969/j.issn.1674-7968.2025.04.009

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摘要麦洼牦牛(Bos grunniens)是青藏高原型牦牛的地方良种,前期研究表明,麦洼牦牛的毛色与其生产性能相关,可作为育种依据,其灰毛性状与突触融合蛋白17 (syntaxin-17, STX17)基因的表达水平相关,STX17蛋白参与溶酶体和自噬体融合过程,在黑色素的合成、运输及黑色素瘤的生成中起重要作用。为研究STX17基因与麦洼牦牛毛色形成的相关性,本研究选取3岁龄的健康成年雄性黑色和灰色麦洼牦牛各3头,以其心、脾、肺、肾和皮肤组织作为实验材料,通过PCR克隆STX17基因编码区序列,利用在线软件对其进行生物信息学分析,用qRT-PCR技术检测STX17基因在麦洼牦牛不同组织及不同毛色种群中的表达情况。结果表明,麦洼牦牛STX17基因CDS区全长909 bp,编码的蛋白质含302个氨基酸,等电点(pI)理论值为5.82,为酸性蛋白;STX17蛋白无信号肽序列,存在1个靶膜相关的可溶性N-乙基马来酰亚胺敏感因子附着蛋白受体(soluble N-ethylmaleimide-sensitive factor attachment protein receptors, SNARE) 结构域和2个串联的跨膜结构域;亚细胞定位预测STX17蛋白主要存在于细胞质中;核苷酸序列比对显示,麦洼牦牛STX17基因与黄牛(Bos taurus)亲缘性最近,其次是水牛(Bubalis bubalus);STX17蛋白与自噬相关蛋白14 (autophagy-related protein 14, ATG14)、突触体相关蛋白29 (synaptosome associated protein of 29, SNAP29)、囊泡相关膜蛋白7 (vesicle associated membrane protein 7, VAMP7)、囊泡相关膜蛋白8 (vesicle associated membrane protein 8, VAMP8)和突触素同源物(synaptobrevin homolog, YKT6)等蛋白存在相互作用;qRT-PCR结果显示,STX17基因在皮肤组织中表达量最高,且在灰毛麦洼牦牛皮肤中的表达量高于黑毛麦洼牦牛,推测STX17基因与麦洼牦牛灰色和黑色毛色的形成有关。本研究为后续麦洼牦牛毛色的调控机制等研究提供理论依据。

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	江明锋
	刘益丽

关键词 ：牦牛, 突触融合蛋白17 (STX17), 毛色, 生物信息学分析, 组织特异性表达

Abstract：Maiwa yak (Bos grunniens) is a local superior seed of Qinghai-Tibet Plateau yak. Previous studies have shown that the coat color of Maiwa yak is related to its production performance and can be used as the basis for breeding. Previous research has shown that the grey coat trait in Maiwa yak is associated with the expression level of the syntaxin-17 (STX17) gene. The STX17 protein is involved in the fusion process of lysosomes and autophagosomes, playing an important role in the synthesis, transport of melanin, and the development of melanoma. To investigate the correlation between the STX17 gene and coat color formation in Maiwa yak, 3 healthy adult male black and grey Maiwa yaks aged 3 years were selected in this study, and their heart, spleen, lung, kidney and skin tissues were selected as experimental materials. The CDS sequence of the STX17 gene were cloned by PCR, and bioinformatics analysis was carried out by using on line software, the expression of the STX17 gene was detected in different tissues and coat populations of Maiwa yak by qRT-PCR. The results showed that the CDS region of the STX17 gene in Maiwa yak was 909 bp, encodimg protein contained 302 amino acids with a theoretical value of isoelectric point (pI) of 5.82, which was an acidic protein, the STX17 protein did not have a signal peptide sequence, and there was 1 target membrane-associated soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) domain and 2 tandem transmembrane domains; Subcellular localisation predicted that the STX17 protein mainly existed in the cytoplasm. The nucleotide sequence comparison showed that the STX17 gene in Maiwa yak was most closely related to Bos taurus, followed by Bubalis bubalus. The STX17 protein was closely related to autophagy-related protein 14 (ATG14), synaptosome associated protein of 29 (SNAP29), vesicle associated membrane protein 7 (VAMP7), vesicle associated membrane protein 8 (VAMP8), synaptobrevin homolog (YKT6) and other proteins. The qRT-PCR results showed that the STX17 gene had the highest expression in skin tissues, and the expression in the skin of grey Maiwa yak was higher than black Maiwa yak, it was hypothesized that the STX17 gene was related to the formation of grey and black coat color in Maiwa yak. This study provides theoretical basis for the subsequent studies on the regulatory mechanism of coat color in Maiwa yak.

Key words： Yak Syntaxin-17 (STX17) Coat color Bioinformatics analysis Tissue-specific expression

收稿日期: 2024-07-26

中图分类号： S823.85

基金资助:那曲市科技局重点研发计划项目(NQKJ-2023-07);西南民族大学中央高校基本科研业务费专项资金(ZYN2024062);四川省区域创新合作项目(2023YFQ0076)

通讯作者: ** mingfengjiang@vip.sina.com;yililiunum@163.com

作者简介: * 同等贡献作者

引用本文:

石琳, 郭子玄, 李小伟, 马士龙, 何小强, 江明锋, 刘益丽. 麦洼牦牛毛色候选基因STX17的克隆及组织表达分析[J]. 农业生物技术学报, 2025, 33(4): 802-813.
SHI Lin, GUO Zi-Xuan, LI Xiao-Wei, MA Shi-Long, HE Xiao-Qiang, JIANG Ming-Feng, LIU Yi-Li. Cloning and Tissue Expression Analysis of Coat Color Candidate Gene STX17 in Maiwa Yak (Bos grunniens). 农业生物技术学报, 2025, 33(4): 802-813.

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https://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2025.04.009 或 https://journal05.magtech.org.cn/Jwk_ny/CN/Y2025/V33/I4/802

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