摘要
旨在克隆牦牛解偶联蛋白1(UCP1)基因序列,并对其进行生物信息学分析,进一步研究该基因在牦牛各组织的表达规律.以牦牛为实验对象,利用RT-PCR克隆得到牦牛UCP1基因全长序列,生物信息学分析牦牛CDS区,qPCR检测UCP1基因在牦牛不同组织中的表达模式.结果表明,UCP1基因全长为954 bp(GenBank No.MW345537),其中开放阅读框(Open reading frame,ORF)全长为942 bp,编码313个氨基酸.蛋白分子特性预测UCP1蛋白为不稳定疏水性蛋白,存在跨膜结构域以及多个磷酸化位点.二级结构和三级结构预测显示,UCP1蛋白由无规则卷曲、α螺旋和延伸链三者交替出现.牦牛UCP1基因的核苷酸和氨基酸序列与普通牛的相应序列相似度最高,同源性分别为97.63%和96.76%;与斑马鱼的核苷酸序列和氨基酸序列相似度最低,分别为62.58%和64.29%.组织表达分析发现UCP1在牦牛心、肝、脾、肺、肾、脂肪、背肌、半腱肌中均有表达,但在肾中的表达量最高,肾与其他组织之间差异显著(P<0.05).不同品种牦牛UCP1基因的差异表达分析发现,其在麦洼牦牛背最长肌、肝中的表达量均显著高于金川牦牛(P<0.05和P<0.0001).这些结果为丰富牦牛UCP1基因的功能研究提供了参考依据.
The aim of this study was to clone the yak uncoupling proteins 1(UCP1)gene sequence,conduct bioinformatics analysis,and further elucidate its expression pattern in various tissues.The full length sequence of yak UCP1 gene was cloned by RT-PCR,the CDS region of UCP1 gene of yak was analyzed by bioinformatics,and the expression of UCP1 gene in different yak tissues was detected by qPCR.The results showed that the full length of UCP1 gene was 954 bp(GenBank No.MW345537),and open reading frame(ORF)of UCP1 gene was 942 bp,encoding 313 amino acids.The prediction showed that UCP1 was an unstable hydrophobic protein with transmembrane domain and multiple phosphorylation sites.The secondary structure and tertiary structure predicted that UCP1 protein included irregular curl,α-helix,and extension chain domains.The nucleotide and amino acid sequence comparison of UCP1 gene showed that the similarity of nucleotide and amino acid sequence was highest between yak and cattle(97.63%and 96.76%,respectively).While the similarity was lowest between yak and zebrafish(62.58%of nucleotide and 64.29%of amino acid).Interestingly,qPCR showed that UCP1 expressed in yak heart,liver,spleen,lung,kidney,fat,longissimus muscle and semitendinosus muscle,but the expression level was highest in the kidney(P<0.05).The mRNAlevel of UCP1 in Maiwa yak longissimus muscle and liver was significantly higher than that of Jinchuan yak(P<0.05 andP<0.0001).These results provided the basic data for functional research of UCP1 gene in yaks.
作者
赵丹
熊燕
华永琳
岳永起
郭玉
熊显荣
字向东
殷实
李键
ZHAO Dan;XIONG Yan;HUA Yong-lin;YUE Yong-qi;GUO Yu;XIONG Xian-rong;YIN Shi;LI Jian(School of Animal&Veterinary Sciences,Southwest Minzu University,Chengdu 610041,China;Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization of Ministry of Education,Southwest Minzu University,Chengdu 610041,China;Key Laboratory of Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Utilization of Sichuan Province,Southwest Minzu University,Chengdu 610041,China)
出处
《西南民族大学学报（自然科学版）》
CAS
2021年第2期139-148,共10页
Journal of Southwest Minzu University(Natural Science Edition)
基金
四川省科技计划项目(2019YJ0258)
国家自然科学基金项目(31902154)。
