莆田黑鸭PMEL17基因的克隆、组织表达与多态性分析  

Cloning,Tissue Expression and Polymorphism Analysis of PMEL17 Gene in Putian Black Duck

作　　者：潘家华 周劢 陈巧慧 林进发 麦丽君 李潇凡 肖天放[1] 林瑞意[1] PAN Jiahua;ZHOU Mai;CHEN Qiaohui;LIN Jinfa;MAI Lijun;LI Xiaofan;XIAO Tianfang;LIN Ruiyi(College of Animal Science(College of Bee Science),Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002;Quanzhou Chengxin Agriculture and Animal Husbandry Development Company,Quanzhou,Fujian 362100)

机构地区：[1]福建农林大学动物科学学院(蜂学学院),福建福州350002 [2]福建省泉州市诚信农牧发展有限公司,福建泉州362100

出　　处：《中国家禽》2020年第12期19-24,共6页China Poultry

基　　金：国家自然科学基金项目(31702109);福建农林大学动物科学学院扶持基金(2018DKOO1);泉州市农业领域科技项目(2018N014)。

摘　　要：为研究莆田黑鸭前黑素小体蛋白(PMEL17)基因序列特征,分析其在不同羽色鸭毛囊及莆田黑鸭不同组织中的表达情况,以莆田黑鸭毛囊组织为材料,利用cDNA克隆技术获得PEML17基因编码区全长序列并进行生物信息学分析;利用荧光定量PCR技术检测PMEL17基因在毛囊、心脏、肝脏、脾脏、肺脏、肌胃、肾脏、大脑、胸肌、腿肌和皮肤组织中的表达情况。结果表明,莆田黑鸭PMEL17基因编码区全长1917 bp,其同源性与鸡最近,与其它物种变异性较大;PMEL17基因编码638个氨基酸,二级结构以无规则卷曲和延伸链为主,预测为稳定的疏水性蛋白,存在1个跨膜区;PMEL17基因在黑羽毛囊中表达,在白羽毛囊中不表达,莆田黑鸭各组织中仅在毛囊和皮肤表达,其它组织几乎不表达;在第6外显子处发现2个SNP位点,导致天冬氨酸突变成丙氨酸。试验结果为进一步研究PMEL17基因在鸭羽色形成中的作用奠定基础,也为莆田黑鸭种质特性研究提供数据。The objective of this study was to analyze the duck PMEL17(premelanosome prolein)gene sequence characteristics,explore its expression in feather bulbs with different plumage colors(Putian Black,Putian White,Liancheng White duck)and different tissues from Putian Black duck.Feather bulb tssue RNA from Putian Black duck was used as a template,and the complete CDs region of duck PMEL17 gene was obtained by cDNA cloning technology and bioinformatics analysis was also conducted.The expression of PMEL17 in heart,liver,spleen,lung,kidney,brain.pectoralis,leg muscle,gizzard,skin,feather bulb tissues from Putian Black duck were detected by fluorescence quantitative PCR.The resuls showed that the full-length of the duck PMEL17 gene codling region was 1917 bp,which had high homology to chicken and great divergence with other species.PMEL17 gene encoded 638 amino acids,and it was predicted to express a stable and hydrophobic protein with one transmembrane region.lts secondary stnucture consists of extended strand and random coil.mRNA expression of PMEL17 was not observed in white feather bulbs.PMEL17 gene was only expressed in skin and feather bulb of Putian Black duck.Two polymorphic sites 145(A/C),146(T/A)were found in the exon 6 of PMELl7 gene,which causes changes in amino acids(D/A).These results laid a foundation for further study of the function of PMELI7 gene in the duck feather color formation and genetic characteristics of Putian Black duck.

关 键 词：莆田黑鸭 PMEL17基因 基因克隆 组织表达 多态性 

分 类 号：S834[农业科学—畜牧学]