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当前位置: 首页 > 文章 > 连城白鸭MITF基因的cDNA克隆、组织表达与分析 福建农业学报 2018 (12) 1292-1300
Position: Home > Articles > cDNA Cloning,Tissue Expression Analysis of MITF Gene in Liancheng White Ducks Fujian Journal of Agricultural Sciences 2018 (12) 1292-1300

连城白鸭MITF基因的cDNA克隆、组织表达与分析

作  者：
辛清武;刘艺展;朱志明;李丽;章琳俐;缪中纬;郑嫩珠
单  位：
福建农林大学动物科学学院;福建省农业科学院畜牧兽医研究所
关键词：
连城白鸭;小眼畸形相关转录因子;序列分析;基因表达
摘  要：
对连城白鸭刺参小眼畸形相关转录因子(microphthalmia-associtated transcription factor,MITF)的序列进行克隆及序列分析,预测MITF蛋白的二、三级结构,并利用RT-PCR技术检测MITF基因在连城白鸭不同组织中的表达差异,为深入研究连城白鸭体内黑色素沉积的内在调控机制奠定理论基础。结果显示,连城白鸭MITF基因长度为1 323bp (登录号:MG516570),包括一个完整的CDS区(34～1 323bp),编码429个氨基酸;与绿头鸭氨基酸同源性100%,与原鸡等鸟类的同源性达95%以上,而与绵羊等哺乳动物的亲缘关系较远。对MITF蛋白的二级结构预测显示,无规则卷曲及α螺旋是形成其二级结构的主要结构元件;连城白鸭MITF蛋白三级结构为一个环状结构间隔开两个螺旋,从而形成MITF蛋白的基础螺旋-环-螺旋-拉链结构。荧光定量PCR分析显示,连城白鸭皮肤组织中的MITF基因mRNA表达量最高,极显著(P<0.01)高于其他各组织;各组织表达量依次为:皮肤>肾脏>肌胃>肝脏>肌肉。综上所述,MITF基因在连城白鸭组织中的表达具有组织特异性,为深入探究MITF基因能否作为调控连城白鸭黑色素沉积的主效候选基因及连城白鸭黑色素性状的辅助选择提供理论依据。
译  名：
cDNA Cloning,Tissue Expression Analysis of MITF Gene in Liancheng White Ducks
作  者：
XIN Qing-wu;LIU Yi-zhan;ZHU Zhi-ming;LI Li;ZHANG Lin-li;MIAO Zhong-wei;ZHENG Nen-zhu;Institute of Animal Science and Veterinary Medicine,Fujian Academy of Agricultural Sciences;College of Animal Science,Fujian Agriculture and Forestry University;
单  位：
XIN Qing-wu%LIU Yi-zhan%ZHU Zhi-ming%LI Li%ZHANG Lin-li%MIAO Zhong-wei%ZHENG Nen-zhu%Institute of Animal Science and Veterinary Medicine,Fujian Academy of Agricultural Sciences%College of Animal Science,Fujian Agriculture and Forestry University
关键词：
Liancheng white duck;;microphthalmia-associated transcription factor;;sequence analysis;;gene expression
摘  要：
The cloning,tissue expression and structural analysis of Microphthalmia-associated transcription factor(MITF)gene were conducted in Liancheng white duck,which would provide a theoretical basis for the further research of melanin deposition.The MITF gene from the skin of female ducks was cloned through nested-PCR,and the mRNA expression detected in tissues by qRT-PCR.It was shown that the MITF cDNA was 1 323 bp in length(GenBank No.MG516570)with a complete CDS region(34-1 323bp)encoding 429 amino acids.The amino acid sequence of MITF was 100% homologous to that of mallard and more than 95% to that of Gallus gallus,but farther from sheep and other mammals.The predicted secondary structure of the protein was mainly random coil andα-helix,while the circular tertiary structure contained two separate helices to form the helix-loop-helix-chain structure.The qRT-PCR results showed that the mRNA expression of MITF was significantly higher in the skin than the other organs(P<0.01)with the relative expressions in the order of skin>kidney>gizzard>liver>muscle.It appeared that the expression of MITF gene was tissue-specific in Liancheng white ducks.The information would aid in the utilization of MITFgene for studies on the melanin deposition and trait selection of the duck.

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