Differential expression of CYB5A in Chinese and European pig breeds due to genetic variations in the promoter region
Differential expression of CYB5A in Chinese and European pig breeds due to genetic variations in the promoter region
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由于启动子区域的遗传变异，CYB5A在中国和欧洲猪品种中的差异表达
DOI：
10.1111/age.12257
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发表时间：
2015-02-01
期刊：
影响因子：
2.4
通讯作者：
Fang, M. Y.
中科院分区：
文献类型：
--
作者：
Bai, Y.;Zhang, J. B.;Fang, M. Y.
细胞色素b5(Cyb5A)是一种重要的电子传递蛋白，在许多不同的生物体中都有同源物。在猪中，CyB5A与野猪的污染有关，因为它在雄酮生物合成中发挥作用。为了研究猪细胞色素B5A基因在猪种中的表达差异，对中国猪和欧洲猪的细胞色素5A基因启动子区域的遗传变异进行了研究。在猪CyB5A启动子区域发现了3个单核苷酸多态(NC_010443.4：g.165901487delG、g.165901767T>；C和g.165902078C>；T)。这些SNPs在中国猪和欧洲猪中的出现频率不同。中国猪主要为单倍型B(记为Delg-C-T：CyB5基因165901487位点为G缺失，165901767位为C位，165902078位为T位)，与欧洲猪相似，但黎查黑猪主要为单倍型A(记为G-T-C：165901487位点为G，165901767位为T，165902078位为C)。肝组织定量聚合酶链式反应结果显示，单倍型B型个体的CyB5A基因表达水平高于单倍型A型个体。体外细胞转染实验也证实了这一点，单倍型B型个体的报告活性高于单倍型A型个体。计算机分析预测，MYC相关的锌指蛋白(MAZ)是一种位于165901767位的潜在转录因子。电泳迁移率改变分析表明，该多态性影响转录因子与CyB5A启动子的稳定结合，进而影响该基因的表达水平。因此，猪CyB5A启动子区域的这种变异可能解释了中国和欧洲猪品种雄酮积累的差异，并可能被证明是区分不同猪品种来源的有用的遗传标记。
Cytochrome b5 (CYB5A) is an important electron transfer protein with homologues in a number of different organisms. In pigs, CYB5A is related to boar taint because of its role in androstenone biosynthesis. To determine the variety of CYB5A expression in pig breeds, genetic variations in the porcine CYB5A promoter region in both Chinese and European pig breeds were examined. Three single nucleotide polymorphisms (NC_010443.4:g.165901487delG, g.165901767T>C and g.165902078C>T) were identified in the porcine CYB5A promoter region. These SNPs occurred in different frequencies in Chinese and European pigs. Chinese pigs were primarily haplotype B (denoted as delG-C-T: the position of nt 165901487 of the CYB5 gene is a G deletion, nt 165901767 is C and nt 165902078 is T), except for Licha black pigs, which were primarily haplotype A (denoted as G-T-C: nt 165901487 is G, nt 165901767 is T and nt 165902078 is C), similar to European pigs. Quantitative PCR data from liver tissues demonstrated that haplotype B individuals had higher CYB5A expression than did those with haplotype A. This was confirmed by in vitro cell transfection assays, in which haplotype B individuals had higher reporter activity than did those with haplotype A. In silico analysis predicted that Myc-associated zinc-finger protein (MAZ) is a potential transcription factor at position 165901767. Electrophoretic mobility shift assays showed this polymorphism affects the stable binding of transcription factors to the CYB5A promoter, which in turn affects the expression levels of this gene. Therefore, this variation of the porcine CYB5A promoter region may explain the differences in androstenone accumulation between Chinese and European pig breeds and may also prove useful as a genetic marker to distinguish the origin of different pig breeds.