广西德保猪MC1R基因的克隆与分析

Cloning and Analysis MC1R Gene of Guangxi Debao Pig

近年来含有天然黑色素的动植物黑色食品得到青睐,为探讨广西地方品种德保猪全黑毛色形成的分子机制,本文克隆了广西德保猪MC1R基因并进行了系统生物信息学分析。根据NCBI已公布的猪MC1R基因序列(GI:347618788)设计特异性引物,以从德保猪血液中提取的基因组DNA为模板,通过PCR扩增克隆获得长1 519 bp的MC1R基因序列片段。多重序列比较结果显示克隆片段与已公布的猪、牛、人、狗、绵羊、小鼠和鸡的相似性依次分别为99%、87%、86%、85%、84%、81%和77%,表明MC1R基因在不同哺乳动物间具有较高的保守型。德保猪MC1R蛋白结构预测结果显示,其理论分子质量为34.65 ku,等电点为8.70,为弱碱性蛋白,N-末端不存在信号肽,定位于细胞质膜,具有1个低复杂度结构域和7个跨膜结构域,具有典型的细胞膜受体蛋白结构。多项研究报道表明MC1R基因是动物毛皮颜色重要的决定基因,德保猪MC1R基因克隆为揭示其全黑色形成的分子机制奠定了较好的实验基础。

In recent years, black food contains natural melanin of plants and animals were paid increasing attention. To explore the molecular mechanism of black coat color of Debao pig, MC1R gene was cloned and bioinformatically analyzed. A pair of primers was designed according to pig MC1R gene sequence （GI： 347618788）. And a 1 519 bp fragment were amplified from the genome DNA purified from the Debao pig blood. The sequence multi-aligned results showed that its similarity with Sus scrofa, Bos taurus, Homo sapiens, Canis lupus, Ovis aries and Mus musculus was up to 99%, 87%, 86%, 85%, 84%, 81% and 77%, respectively, which suggested that MC1R gene was high conservative among mammals. The further protein structure analysis results showed that the protein weight 34.65 ku and isoelectric point was 8.7, without N-terminal signal peptide, localized in cell membrane with a low complexity domain and typical seven transmembrane domains. A few of reports revealed that MC1R gene was an important decision factor affected animal fur color, the study provided a good basis to reveal the whole black color forming molecular mechanism of Debao pig.