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牦牛FABP3基因克隆及组织表达谱分析
戴荣凤1,2, 黄 纯1,2, 扎 老,等3
1.中国农业科学院 兰州畜牧与兽药研究所 甘肃省牦牛繁育工程重点实验室;2.农业农村部 青藏高原畜禽遗传资源育种重点实验室;3.合作市佐盖多玛乡畜牧站
摘要:
【目的】 分析牦牛心肌脂肪酸结合蛋白3(FABP3)基因的结构和功能,并检测其在成年牦牛和胎牛中的表达水平,为探究该基因在牦牛育种中的生物学功能提供理论参考。【方法】以美仁牦牛的心脏组织为试验材料,PCR扩增FABP3基因,对得到的编码区序列(CDS)进行生物信息学分析;利用实时荧光定量PCR(RT-qPCR),检测FABP3基因在成年牦牛和胎牛心脏、肝脏、脾脏、肺脏、肾脏及肌肉组织的表达水平。【结果】牦牛FABP3基因编码区序列长度为402 bp,编码133个氨基酸;蛋白质的高级结构主要是β 转角和延伸链;牦牛FABP3蛋白不存在跨膜区域和信号肽,属于具有一定亲水性的稳定蛋白;牦牛FABP3基因CDS区核苷酸及其编码氨基酸序列的同源性分析显示,FABP3基因在牛属动物中具有一定的保守性;系统进化树分析表明,牦牛与野牦牛和瘤牛的亲缘关系最近,与鸡的亲缘关系最远。RT-qPCR结果显示,FABP3基因在成年牦牛和胎牛的心脏、肝脏、脾脏、肺脏、肾脏及肌肉组织中均有表达,但在胎牛肝脏、脾脏、肾脏和肌肉中的表达水平显著或极显著高于成年牦牛,在成年牦牛肺脏中的表达水平极显著高于胎牛。【结论】克隆了牦牛FABP3基因,探究了FABP3基因在牦牛中的组织表达规律,为进一步研究该基因在牦牛脂肪沉积中的作用提供了基础数据。
关键词:  美仁牦牛  FABP3  基因克隆  生物信息学  组织表达谱
DOI:
分类号:
基金项目:国家重点研发计划项目(2021YFD1600200);合作市牦牛种质提升与提质增效项目;甘肃省科技计划项目(20JR5-RA580);甘肃省科技重大项目(21ZD10NA001,GZGG-2021-1);现代农业(肉牛牦牛)产业技术体系建设专项(CARS-37);中国农业科学院科技创新工程项目(25-LZIHPS-01)
Cloning and tissue expression profile analysis of FABP3 gene in Bos grunniens
DAI Rongfeng,HUANG Chun,ZHA Lao,et al
Abstract:
【Objective】The structure and function of yak heart fatty acid binding protein 3 (FABP3) gene were analyzed and its expression levels in adult and fetal yak were detected to provide references for exploring biological functions of the gene in yak breeding.【Method】The FABP3 gene was amplified by PCR from heart tissue of Meiren yak,and bioinformatics analysis was carried out after the CDS region was obtained.The expression levels of FABP3 in heart,liver,spleen,lung,kidney,and muscle tissues of adult and fetal yak were detected by real-time quantitative PCR (RT-qPCR).【Result】The length of the coding region of FABP3 was 402 bp,encoding 133 amino acids.The advanced structure of the protein included mainly β-turn and extended chain.There was no transmembrane region or signal peptide on yak FABP3 protein,indicating it was a stable protein with certain hydrophilicity.Homology analysis of the nucleotide and amino acid sequences of the CDS region showed that the FABP3 gene was conserved in cattle.According to the phylogenetic tree,yak had the closest genetic relationship with wild yak and zebu,and the farthest genetic relationship with chicken.RT-qPCR results showed that the FABP3 gene was expressed in heart,liver,spleen,lung,kidney,and muscle tissues of adult and fetal yak.The expression in liver,spleen,kidney,and muscle of fetal cattle was significantly or extremely significantly higher than that of adult yak,and the expression level in lung of adult yak was significantly higher than that of fetal yak.【Conclusion】The FABP3 gene of yak was cloned,and its tissue expression pattern was explored.This study provided basic data for further study on the role of FABP3 in fat deposition.
Key words:  Meiren yak  FABP3  gene cloning  bioinformatics  tissue expression profile

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