摘要
【目的】探究FABP4和FABP5基因对牦牛脂肪酸的调控机制,提升牦牛肉品质。【方法】采用RT-PCR扩增牦牛FABP4和FABP5基因CDS序列,利用在线软件进行生物信息学分析,使用MEGA7.0构建系统发育进化树分析其同源性。采用实时荧光定量PCR检测FABP4和FABP5基因在心脏、肝脏、肺脏、肾脏、脂肪、肌肉和十二指肠中的表达水平,探究其在不同组织中的表达差异。【结果】牦牛FABP4、FABP5基因CDS序列全长分别为393和408 bp,分别编码130和135个氨基酸,基于FABP4和FABP5氨基酸序列相似性构建的系统进化树均显示,牦牛与美洲野牛的亲缘关系最近,其次是牛,与鸡的亲缘关系较远。FABP4和FABP5蛋白二级结构、三级结构以无规则卷曲和延伸链为主,无跨膜结构域和信号肽,推测其不是分泌蛋白,为稳定的亲水性蛋白。牦牛FABP4、FABP5氨基酸序列分别存在16和17个磷酸化位点,均以苏氨酸磷酸化位点为主;牦牛FABP4和FABP5蛋白均与PPARG、GOT2间存在互作关系,且其相互间存在互作关系。KEGG富集结果发现,FABP4和FABP5共同富集在PPAR信号通路上。FABP4和FABP5基因在牦牛各组织中均有不同程度的表达,其中在脂肪中的相对表达量最高。【结论】克隆了牦牛FABP4和FABP5基因,明确了其分子特征。牦牛FABP4和FABP5基因在脂肪组织中高表达,与脂肪的沉积和代谢密切相关。
【Objective】This research aimed to investigate the mechanism of FABP4 and FABP5 genes on the regulation of fatty acids in yak,to improve the quality of yak meat and to accelerate the promotion of yak meat.【Method】The sequences of protein coding region(CDS)of yak FABP4 and FABP5 genes were amplified by RT-PCR.Bioinformatic analysis was conducted with online software,and their homology was analyzed by constructing phylogenetic evolutionary tree with MEGA7.0.Real-time fluorescence quantitative PCR was used to detect the expression of FABP4 and FABP5 genes in heart,liver,lung,kidney,adipose,muscle and duodenum,and to explore the differences in their expression in different tissues.【Result】The full-length CDS sequences of yak FABP4 and FABP5 genes were 393 and 408 bp respectively,encoding 130 and 135 amino acids,respectively.Phylogenetic trees constructed on the basis of similarity of amino acid sequences of FABP4 and FABP5 both showed that yak was most closely related to American bison,followed by cattle,and was less closely related to chicken.The secondary and tertiary structures of the protein were dominated by irregular coils and extended chains.With no transmembrane structural domains or signal peptides,it was presumed that it was not a secreted protein but a stable hydrophilic protein.The amino acid sequences of yak FABP4 and FABP5 contained 16 and 17 phosphorylation sites,respectively,of which threonine phosphorylation sites were dominant.Protein-protein interaction analysis revealed interactions between yak FABP4 and FABP5 proteins with PPARG,GOT2,and each other,and the results of KEGG enrichment showed that FABP4 and FABP5 were co-enriched in the PPAR signaling pathway.FABP4 and FABP5 genes were expressed in different degrees in all tissues of yak,with the highest relative expression in fat tissue.【Conclusion】Yak FABP4 and FABP5 genes were cloned and their molecular characteristics were analyzed.Yak FABP4 and FABP5 genes were highly expressed in adipose tissue and were closely related to fat deposition and metabolism.
作者
张梦帆
洛桑顿珠
喇永福
任稳稳
马晓明
张强
卓玛次仁
尼玛加措
平措占堆
梁春年
ZHANG Mengfan;LUOSANG Dunzhu;LA Yongfu;REN Wenwen;MA Xiaoming;ZHANG Qiang;ZHUOMA Ciren;NIMA Jiacuo;PINGCUO Zhandui;LIANG Chunnian(Key Laboratory of Yak Breeding Engineering in Gansu Province,Lanzhou Institute of Husbandry and Pharmaceutical Science,Chinese Academy of Agricultural Sciences,Lanzhou,Gansu 730050,China;Key Laboratory of Livestock and Poultry Genetics and Breeding on the Qinghai-Tibet Plateau,Ministry of Agriculture and Rural Affairs,Lanzhou,Gansu 730050,China;Institute of Husbandry and Veterinary Research,Xizang Academy of Agricultural and Animal Husbandry Sciences,Lhasa,Xizang 850004,China;Agriculture and Rural Affairs Bureau of Angren County,Rikaze,Xizang 857001,China)
出处
《西北农林科技大学学报(自然科学版)》
北大核心
2025年第7期9-17,共9页
Journal of Northwest A&F University(Natural Science Edition)
基金
国家重点研发计划项目(2022YFD1302101)
西藏自治区区域科技协同创新项目(QYXTZX-RKZ2022-03)
现代肉牛牦牛产业技术体系项目(CARS-37)。
