红褐色乌苏里貉MC1R基因的克隆及序列分析

为了探讨红褐色貉的促黑素细胞激素受体(MC1R)的序列长度、多态性以及与其他物种的同源性。以青岛莱西养殖基地的野生型乌苏里貉8只，红褐色乌苏里貉7只，吉林白貉6只，银黑狐6只为研究对象，用提取貉和银黑狐的毛囊DNA作为模板，根据GenBank已知序列(GenBank：HM852533.1)设计引物，利用克隆测序技术，通过SNP检测，分析MC1R基因在不同毛色乌苏里貉的表达水平和毛色之间相关性以及与其他物种的同源性。结果表明：本实验成功成功获得了红褐色貉MC1R基因序列长度为1329bp，与已知序列同源性98%，13处突变，与野生型乌苏里貉同源性达到99%，8处突变。通过酶切鉴定在MC1R基因约300bp处酶切得到2种基因型AB型和BB型，红褐色貉为AB型，野生型貉为BB型。根据实验结果推测MC1R基因存在红褐色毛色性状相关的功能突变位点，从而导致毛色突变。

Cloning and Sequence Analysis of MC1R Gene in Reddish-brown Wusuli Raccon Dog

To study the sequence length in MC1R gene of Reddish-brown Wusuli Raccon Dog,the polymorphism and homology of the other species.In the experiment for research object,that had 8 wild Wusuli Raccoon Dog,7 reddish-brown Wusuli Raccoon Dog,6 Jilin White Raccoon Dog and 6 silver-black fox from Qingdao Lacy breeding base.By the SNP testing,we analyzed could the correlation of MC1R gene expression level in the Wusuli Raccoon Dog in different colors.As known as NCBI sequence(GenBank:HM852533.1),with the extraction of Raccoon Dog hair follicle DNA as a template,we designed primer,sing for getting MC1R gene which play an important role in mammals melanin cell synthesis melanin process.The results showed that we was successful to get the Reddish-brown Raccoon Dog MC1R gene sequence for 1329 bp,and the known sequence homology 91.47%,18 mutations;with the wild type Wusuli Raccoon Dog homology reached 99%,8 mutations.Through the enzyme cut in MC1R genes,two genes were got enzyme type AB and BB type about 300 bp,Reddish Brown Raccoon Dog for AB,wild type Raccoon Dog for BB type.According to the results of the experiment MC1R genes that there were brown coat of related functions mutations,which could led to coat mutations.