CAJ | 학술논문

以Agouti基因作为牦牛毛色调控的候选基因，探索Agouti基因编码区序列多态性，以期为阐明Agouti与毛色形成的相关性及毛色形成机理奠定基础。根据GenBank已发表序列（ NM_206843）设计1对引物，以天祝白牦牛和黑色被毛甘南牦牛皮肤总RNA为模板，采用RT-PCR技术扩增，成功获得了牦牛Agouti基因编码区序列。采用生物信息学方法预测分析Agouti基因及其编码蛋白的基本理化性质、信号肽、疏水性、二级结构等；采用DNA混合池测序法和直接测序法，检测牦牛Agouti基因编码区的序列变异。结果表明，扩增得到的Agouti基因的编码区是一条长为593 bp的DNA序列。黑色甘南牦牛和天祝白牦牛皮肤的Agouti基因序列相同，并与黄牛基本一致。将牦牛的Agouti基因序列命名为YAK ASIP，并且在NCBI数据库中注册该基因的登录号为KJ630463。该基因含有一个长度为402 bp的开放性阅读框，编码133个氨基酸。 Agouti基因编码蛋白属于亲水性蛋白，有一个明显的信号肽，含有多个磷酸化位点。其二级结构主要以α螺旋和无规则卷曲为主。 Agouti基因编码产物氨基酸邻接系统树表明，牦牛Agouti与黄牛、绵羊等物种的Agouti氨基酸具有高度相似性，并且牦牛Agouti基因编码区中无多态性位点。
이Agouti기인작위모우모색조공적후선기인，탐색Agouti기인편마구서렬다태성，이기위천명Agouti여모색형성적상관성급모색형성궤리전정기출。근거GenBank이발표서렬（ NM_206843）설계1대인물，이천축백모우화흑색피모감남모우피부총RNA위모판，채용RT-PCR기술확증，성공획득료모우Agouti기인편마구서렬。채용생물신식학방법예측분석Agouti기인급기편마단백적기본이화성질、신호태、소수성、이급결구등；채용DNA혼합지측서법화직접측서법，검측모우Agouti기인편마구적서렬변이。결과표명，확증득도적Agouti기인적편마구시일조장위593 bp적DNA서렬。흑색감남모우화천축백모우피부적Agouti기인서렬상동，병여황우기본일치。장모우적Agouti기인서렬명명위YAK ASIP，병차재NCBI수거고중주책해기인적등록호위KJ630463。해기인함유일개장도위402 bp적개방성열독광，편마133개안기산。 Agouti기인편마단백속우친수성단백，유일개명현적신호태，함유다개린산화위점。기이급결구주요이α라선화무규칙권곡위주。 Agouti기인편마산물안기산린접계통수표명，모우Agouti여황우、면양등물충적Agouti안기산구유고도상사성，병차모우Agouti기인편마구중무다태성위점。
In yak,Agouti gene is a potential candidate gene in yak skin′s color formation and regulation ,inves-tigate in Agouti gene sequence polymophism lay the foundation for revealing yak skin and coat color genetic mecha-nisms.The presented study using DNA cloning techniques to obtain yak Agouti gene encoding sequences .Using bioinformatics tools to forecast the protein′s basic physical,chemical properties,hydrophobicity,signal peptide and secondary structure .The polymorphisms of Agouti gene CDS were identificated by DNA pools and direct sequencing in Yak.The results showed that the obtained Agouti was a 593 bp DNA sequence .Gannan black yak Agouti gene was identical with that of Tianzhu white yak .Also,yak Agouti gene was similar with Bos .The yak Agouti gene was named YAK ASIP and registered number in NCBI was KJ 630463 .This gene encode a hydrophilic protein which con-taining 133 amino acids.It contains a clear signal peptide and several phosphorylation sites .The secondary structure of the protein was αhelix secondary structure and random coil .The Agouti and related proteins of Bos grunniens, Bos taurus,Ovis aries are close in the phylogenetic tree ,which showed highly homology .The gene sequence detection showed that the yak Agouti gene was without any polymorphisms .