摘要
试验旨在探究脂联素(adiponectin,ADIPOQ)基因在不同月龄寒泊羊背最长肌组织中的表达规律及生物学特征。采用实时荧光定量PCR方法对3个不同时期(1、7、13月龄)寒泊羊背最长肌组织中ADIPOQ基因mRNA表达水平进行检测,并利用生物信息学方法分析ADIPOQ基因的核苷酸及编码氨基酸序列分子结构特征、蛋白特性及网络互作蛋白。结果显示,ADIPOQ基因在1、7、13月龄寒泊羊背最长肌组织中均有表达,并随着月龄增加呈现增加趋势,无显著性差异(P>0.05)。绵羊ADIPOQ基因核苷酸及氨基酸序列与山羊(98.60%和99.00%)、牛(98.60%和87.00%)的同源性较高。蛋白理化性质分析发现,ADIPOQ蛋白分子式为C1172H1776N314O349S5,分子质量为26.00 ku,理论等电点(pI)为5.88,推测该蛋白为酸性蛋白;ADIPOQ亲水性总平均值(GRAVY)为-0.403,推测该蛋白为亲水性蛋白,不属于跨膜蛋白;信号肽序列为Met1~Ala17,切割位点在Ala17和Arg18之间;在45-101位氨基酸处存在Collagen结构域,101-237位氨基酸处为C1Q保守结构域;二级结构主要以无规则卷曲为主(66.95%)。蛋白互作网络构建分析表明,该蛋白主要与ADIPOR1、ADIPIR2、CDH13和LEP蛋白具有互作关系。本研究结果为进一步深入探讨寒泊羊ADIPOQ基因在肌肉发育过程中的分子生物学功能奠定了基础。
The aim of this study was to investigate the expression and biological characteristics of adiponectin(ADIPOQ)gene in longissimus dorsi muscle of Hanper sheep at different months of age.The ADIPOQ gene mRNA expression was detected in longissimus dorsi muscle of Hanper sheep at 3 different growth periods(1,7 and 13 months old)by Real-time quantitative PCR.The bioinformatics method was used to analyze the nucleotide and amino acid sequences of ADIPOQ gene,protein characteristics and network interaction protein.The results showed that ADIPOQ gene was expressed in longissimus dorsi muscle of Hanper sheep,which showed an increasing trend with the growth of the months age without significant difference(P>0.05).There was higher homology of ADIPOQ gene with Capra hircus(98.60%,99.00%)and Bos taurus(98.60%,87.00%)in nucleotide and amino acid sequences.The results of physical and chemical properties analysis showed that the molecular formula of ADIPOQ protein was C1172H1776N314O349S5,the molecular mass was 26.00 ku,and the theoretical isoelectric point(pI)was 5.88,which suggested that the protein was acidic.The average value of ADIPOQ hydrophilic protein(GRAVY)was-0.403,which indicated that the protein was hydrophilic and didn’t belong to transmembrane protein.In addition,the signal peptide sequence was Met1-Ala17,and the cleavage site was between Ala17 and Arg18.ADIPOQ existed collagen structure domain and C1Q conserved structure domain at position 45-101 and 101-237,respectively.The secondary structure mainly composed of random coil(66.95%).The construction of protein interaction network indicated that ADIPOQ might interact with ADIPOR1,ADIPOR2,CDH13 and LEP.The results provided a theoretical basis for further exploring the molecular biological functions of ADIPOQ gene in the process of muscle development in Hanper sheep.
作者
刘爱菊
赵娟
张鑫
周荣艳
田树军
白莹
陈晓勇
LIU Aiju;ZHAO Juan;ZHANG Xin;ZHOU Rongyan;TIAN Shujun;BAI Ying;CHEN Xiaoyong(College of Animal Science and Technology,Hebei Agricultural University,Baoding071000,China;College of Life Science and Food Engineering,Hebei University of Engineering,Handan 056000,China)
出处
《中国畜牧兽医》
CAS
北大核心
2020年第12期3805-3814,共10页
China Animal Husbandry & Veterinary Medicine
基金
农业农村部政府采购服务项目(16190050)
河北省重点研发计划项目(20326349D)
河北省农业科技成果转化项目(20826603D)
河北农业大学人才引进项目(YJ201938)。