关岭牛FABP3和FABP4基因的分子特征及其组织表达分析  

Molecular characteristics of FABP3 and FABP4 genes and tissue expression analysis of Guanling cattle

作　　者：石鹏飞 阮涌[1] 刘文娇 许家利 孙金魁 熊讯 许厚强[1] SHI Peng-fei;RUAN Yong;LIU Wen-jiao;XU Jia-li;SUN Jin-kui;XIONG Xun;XU Hou-qiang(College of Animal Sciences/Key Laboratory of Animal Genetics,Breeding and Reproduction in the Plateau Mountains Region,Ministry of Education/Key Laboratory of Animal Genetics,Breeding and Reproduction in Guizhou,Guizhou University,Guiyang,Guizhou 550025,China)

机构地区：[1]贵州大学动物科学学院/高原山地动物遗传育种与繁殖教育部重点试验室/贵州省动物遗传育种与繁殖重点试验室,贵州贵阳550025

出　　处：《南方农业学报》2022年第8期2281-2293,共13页Journal of Southern Agriculture

基　　金：国家自然科学基金项目(31571279);教育部促进与美大地区科研合作与高层次人才培养项目([2015]2062号)。

摘　　要：【目的】探究关岭牛心脏型脂肪酸结合蛋白(FABP3)和脂肪型脂肪酸结合蛋白(FABP4)基因的分子特征及其在不同年龄段和不同组织中的表达差异,为揭示牛FABP3和FABP4基因对脂肪酸的调控作用机制提供理论依据。【方法】通过RT-PCR扩增关岭牛FABP3和FABP4基因蛋白编码区(CDS)序列,采用ProtScale、ProtParam、SOPMA、SWISS-MODEL、TMHMM-2.0、SignalP-5.0及NetPhos-3.1等在线软件进行生物信息学分析,同时以实时荧光定量PCR检测FABP3和FABP4基因在3日龄关岭犊牛、24月龄关岭牛(成年)及24月龄杂交牛(关岭牛×利木赞牛)各组织中的表达情况。【结果】关岭牛FABP3、FABP4基因CDS序列全长分别为402和399 bp,对应编码133和132个氨基酸残基,其蛋白二、三级结构以无规则卷曲和延伸链为主,均无跨膜结构域和信号肽,且为稳定的亲水性蛋白。关岭牛FABP3、FABP4氨基酸序列分别存在26和21个磷酸化位点,其中又以苏氨酸磷酸化位点为主;关岭牛FABP3和FABP4蛋白与FABP1、FABP6、过氧化物酶体增殖物激活受体γ(PPARγ)、脂蛋白脂肪酶(LPL)及激素敏感脂肪酶(LIPE)等存在互作关系。基于FABP3和FABP4氨基酸序列相似性构建的系统发育进化树均显示,关岭牛与牦牛的亲缘关系最近,其次是绵羊,与鸡和斑马鱼的亲缘关系较远。FABP3和FABP4基因在关岭牛和杂交牛各组织中均有不同程度的表达,其中,FABP3基因以在心脏中的相对表达量最高,且在成年牛中的相对表达量极显著高于犊牛和杂交牛(P<0.01,下同);FABP4基因以在脂肪中的相对表达最高,且在犊牛中的相对表达量极显著高于成年牛。【结论】关岭牛FABP3和FABP4基因具有较高的遗传保守性,且以在心脏和脂肪中的表达水平较高,与脂肪酸代谢密切相关。【Objective】To investigate molecular characteristics of heart-type fatty acid binding protein(FABP3) and fatty acid binding protein(FABP4)genes and their expression in different age groups and different tissues in Guanling cattle,so as to provide a theoretical basis for revealing mechanism of fatty acid regulation by FABP3 and FABP4 genes in cattle.【Method】The protein coding region(CDS) sequences of FABP3 and FABP4 genes in Guanling cattle were amplified by RT-PCR,and bioinformatics analysis was performed using online software such as ProtScale,ProtParam,SOPMA,SWISS-MODEL,TMHMM-2.0,SignalP-5.0 and NetPhos-3.1.The expression of FABP3 and FABP4 genes in various tissues of 3-day-old Guanyin calves,24-month-old Guanyin cattle(adults)and 24-month-old crossbred cattle(Guanyin cattle × Limousin cattle)was examined by quantitative real-time PCR(qRT-PCR).【Result】The CDS sequences of Guanling cattle FABP3 and FABP4 genes were 402 and 399 bp long,encoding 133 and 132 amino acid residues respectively.Their protein secondary and tertiary structures were mainly irregularly coiled and extended chains without transmembrane structural domains or signal peptide shear sites,and they were stable hydrophilic proteins.FABP3 and FABP4 amino acid sequences had 26 and 21 phosphorylation sites,respectively,among which threonine was more widely distributed;the proteins of FABP3 and FABP4 had interactions with FABP1,FABP6,peroxisome proliferator-activated receptor γ(PPARγ),lipoprotein lipase(LPL)and hormone-sensitive lipase(LIPE).Phylogenetic trees based on amino acid sequence similarity between FABP3 and FABP4 showed that Guanling cattle were most closely related to yaks,followed by sheep,and more distantly related to chickens and zebrafish.FABP3 and FABP4 genes were differentially expressed in all tissues of Guanling and crossbred cattle.The relative expression of FABP3 gene was the highest in the heart and was significantly higher in adult Guanling cattle than in calves and crossbred cattle(P<0.01,the same below);the relati

关 键 词：关岭牛 FABP3基因 FABP4基因 分子特征 组织表达 脂肪酸代谢 

分 类 号：S823.81[农业科学—畜牧学]