瓯江彩鲤体色调控相关因子MC1R的克隆与表达分析  

Cloning and expression analysis of color-related regulation factor MC1R in Oujiang color common carp

作　　者：胡建尊[1] 李康乐[1] 项松平 王剑 朱丽艳[1] 王成辉[1] 

机构地区：[1]上海海洋大学农业部淡水水产种质资源重点实验室,上海201306 [2]浙江龙泉省级瓯江彩鲤良种场,浙江龙泉323700

出　　处：《上海海洋大学学报》2013年第4期518-523,共6页Journal of Shanghai Ocean University

基　　金：国家公益性行业[农业]科研专项(200903045);上海市科学技术委员会项目(123919N1100);浙江省科学技术委员会项目(2010C32088)

摘　　要：黑素皮质素1受体(melanocortin 1 receptor,MC1R)是动物黑色素合成通路中的关键基因之一,对动物的体色和毛色有重要影响。瓯江彩鲤(Cyprinus carpio var.color)具有5种基本体色类型("全红"、"粉玉"、"大花"、"粉花"和"麻花"),其中后3种带有黑色斑点或斑块,是研究动物黑色素合成机制的良好材料之一。克隆了瓯江彩鲤的MC1R基因,并进行了"全红"、"大花"、"粉玉"、"粉花"4种体色间的表达差异分析。研究发现,瓯江彩鲤的MC1R基因的全长cDNA序列为1 914 bp,包括637 bp 5'-UTR(非转录区)、311 bp 3'-UTR及966 bp ORF(开放阅读框)。该基因由一个外显子编码(321个氨基酸),有7个跨膜结构域。瓯江彩鲤与鲫的编码区核苷酸同源性达98%,与斑马鱼达93%;不同体色间的编码区只存在一个核苷酸无义突变(C/G),因而氨基酸序列完全相同。qRT-PCR表明,MC1R基因在眼睛的表达量显著高于皮肤、肌肉、鳃、肾脏、鳔、心、肝等组织(P<0.05);但在4种体色间不存在显著的表达差异(P>0.05)。研究结果表明:瓯江彩鲤体色类型中的黑色斑块不是由MC1R基因直接决定,还有待对其它体色相关基因或调控因子的研究。Melanocortin 1 receptor （MC1R）, one of the key genes in animal melanin synthesis pathway, plays an important role in determining animal body color and coat color. Oujiang color common carp （ Cyprinus carpio var. color） , characterized as five kinds of body color patterns with black patches （ e. g. ＂whole red＂, ＂whole red with big black patch＂, ＂whole red with scattered black spots＂ ,＂ whole white＂ and ＂whole white with big black patch＂） , has been considered as a good material or model in studying animal melanin synthesis mechanism. In this paper, MCIR gene was cloned and sequenced, and its expression profile in the four body color patterns （ ＂whole red＂, ＂whole red with big black patch＂, ＂whole white＂ and ＂whole white with big black patch＂ ） and tissues were also analyzed in Oujiang color common carp. The result showed that the full-length cDNA of MC1R was 1 914 bp, containing a 5＇un-translated region （UTR） of 637 bp, a 3＇-UTR of 311 bp and an open reading frame （ORF） of 966 bp. This gene has only one exon encoding 321 amino acids and predicting 7 transmembrane domains. There were 98% nucleotide similarity with Carassius auratus and 93 % similarity with Danio rerio. However, Only one nucleotide nonsense mutation （ C/ G） in the coding region was found among different color patterns, indicating the same amino acid sequence among them. The result of qRT-PCR indicated that the expression of MC1R in eyes was significantly higher （P 〈0.05） than that in skin, muscles, gills, kidney, bladder, heart and liver. It was surprising that there was no significant expression difference among the different color patterns （ P 〉 0.05 ）. This finding showed that MC1R gene is not a determinant factor in controlling color patterns of Oujiang color common carp, and it is necessary to conduct further studies in other color-related genes or factors.

关 键 词：瓯江彩鲤 黑斑体色 MC1R 表达差异 相关性 

分 类 号：S917[农业科学—水产科学]