羊驼MC1R基因克隆及其在不同毛色个体中表达水平研究

Cloning and Expression Level Analysis of MC1R Gene in Alpacas with Different Coat Color

为了获取羊驼MC1R基因序列,探索MC1R基因表达水平与羊驼毛色之间的相关性,根据GenBank已发表序列(EU1358800)设计1对引物,以羊驼皮肤总RNA为模板,采用RT-PCR技术扩增并成功获得了中华白色羊驼MC1R基因cDNA序列,长度为1081 bp,编码317个氨基酸,与已发表序列进行对比,同源性为99%,发现7处突变。根据所克隆序列设计引物,采用实时荧光定量PCR技术(QRT-PCR),分析MC1R基因在白色和棕色羊驼皮肤中的基因表达量,并对结果进行统计分析。结果表明:经过内参基因校正后,棕色羊驼皮肤中MC1R基因的相对表达量是白色羊驼相对表达量的4.32倍(P<0.01)。MC1R基因表达量的差异与羊驼毛色表型之间可能存在一定的相关性。

In order to obtain the sequence of alpaca MC1R gene and explore the relationship be- tween the expression level of MC1R gene and alpaca coat color, according to the sequence in Gen- Bank （EU1358800）, a pair of specific primer was designed to amplify the cDNA sequence of MC1R using RT-PCR technique. The MC1R gene sequence of white alpaca was cloned successful- ly and sequence analysis revealed that the MC1R gene, encoding 317 amino acids, was 1 081 bp in length. Compared with the published sequence in GenBank, the sequence identity was 99%. Also, 7 mutations were found within the gene. Further, a pair of primer was designed according to the sequence obtained. The relative expression level of MC1R gene in different coat color alpaca was analyzed using QRT-PCR technique and SPSS 13.0 software. The relative gene expression level of MC1R in brown alpaca was 4.32 times than that in white alpaca after normalization with GAPDH gene（P〈0.01）. It was indicated that the gene expression level of alpaca MC1R may be related to the coat color.