摘要
根据GenBank中公布的雪貂（登录号：EF405957）、大熊猫（登录号：XM_002930310）、家犬（登录号：CFU42219）及猫（登录号：AY959314）的酪氨酸酶（tyrosinase,TYR）基因DNA序列保守区域,设计1对特异性引物,利用PCR及克隆技术测定了短毛黑水貂、红眼白水貂和米黄色水貂的TYR基因外显子1部分序列,并对该序列进行了BLAST鉴定及变异位点分析,基于该基因编码氨基酸序列构建了16个物种的系统进化树。结果表明,测定的3种毛色水貂TYR基因序列长度均为796bp,包括795bp的外显子1和1bp 5′UTR,共检测到5个单一突变位点：g.A34G、g.T138A、g.T248C、g.A545G和g.C765A,包含4个错义突变：g.A34G（p.Ser12Gly）、g.T248C（p.Val83Ala）、g.A545G（p.Tyr182Cys）和g.C765A（p.His255Gln）,1个移码突变分别出现在短毛黑水貂和红眼白水貂个体间：g.T138A（p.Cys46＊）,获得TYR基因GenBank登录号分别为：短毛黑水貂（KJ198847）、红眼白水貂（KJ658343）和米黄色水貂（KJ152769）。水貂与雪貂、大熊猫、家犬、猫、家兔、猪、羊驼、山羊、绵羊、牛、人、猕猴、黑猩猩、原鸡和日本鹌鹑的TYR基因核苷酸序列同源性为72.8%～98.6%,相应氨基酸序列同源性为75.0%～98.1%。TYR基因分子进化树显示水貂与雪貂遗传关系最近,分化时间约为1000万年前,且与原鸡和日本鹌鹑亲缘关系最远。该结果为进一步开展TYR基因多态性与水貂毛色表型的相关性研究奠定了生物信息学基础。
One pair of special primer was designed according to the conservative region of tyrosinase （TRY） gene in Mustela putorius f uro , Ailuropoda melanoleuca , Canis lups familiaris and Fells catus deposited in GenBank database. The partial sequence of exon 1 from Neovison vison （Black）, Neovison vison （Regal White） and Neovison vison （Palomino） mink were obtained by PCR amplification and cloning sequencing. The results showed that the length of TYR gene in mink with three kinds of coat color was 796 bp, comprising partial exon 1（795 bp） and 5′UTR （1 bp）. Five singleton variable sites were detected at position 34 （A/G）, 138 （T/A）, 248（T/C）, 545（A/G） and 765（C/A）, including four missense mutations and one frameshift mutation. The accession numbers were KJ198847（Black）, KJ658343（Regal White） and KJ152769 （Palomino）, respectively. The nucleotide similarity between mink and ferret, giant panda, dog, eat, rabbit, pig, alpaca, goat, sheep, cattle, human, rhesus monkey, chimpanzee, chicken and japanese quail was 72.8% to 98. 6%, and the amino acid similarity was 75.0% to 98.1%. The phylogenetic tree constructed by TYR amino acid indicated that the evolution distance of mink was the most homogeneous to ferret beloning to Mustelidae. This study would lay the bioinformatics foundation for further investigating correlations between polymorphisms of TYR gene and coat color phenotype in mink.
出处
《中国畜牧兽医》
CAS
北大核心
2014年第11期205-212,共8页
China Animal Husbandry & Veterinary Medicine
基金
国家重点基础研究发展计划--973项目(2012CB-722907)
关键词
水貂
毛色变异
酪氨酸酶基因
遗传进化
mink
variation of coat color
tyrosinase gene
genetic evolution
