摘要
本研究旨在克隆草原红牛脂肪酸结合蛋白7基因(FABP7)的CDS区序列并对其进行生物信息学分析,同时检测其在牛各个组织中的mRNA表达水平。利用RT-PCR技术克隆草原红牛FABP7基因CDS区序列,并使用多种生物软件和在线工具进行生物信息学分析,通过qPCR技术检测FABP7基因在各组织间mRNA的表达水平。结果表明:草原红牛FABP7基因CDS区全长399 bp,编码132个氨基酸,蛋白分子量为14.96 ku,理论等电点为5.38,属于亲水性蛋白;通过NCBI-BLAST对比发现,草原红牛与普通牛、羊、猪、人、鼠、鸡的核苷酸序列同源性分别为99%、98%、94%、92%、85%、85%,系统进化树结果发现,草原红牛与普通牛亲缘关系最近,与鸡的亲缘关系最远;FABP7蛋白序列有1个二硫键,14个磷酸化位点,1个N-糖基化位点,不存在信号肽和跨膜区;在蛋白的二级结构和三级结构中发现,FABP7蛋白主要存在2个α-螺旋结构和10条β-折叠,为混合型蛋白。FABP7基因在小肠组织表达量最高,在心脏、脂肪和胃中中度表达。
The aim of this study was to clone the CDS region sequence of Red Steppe cattle fatty-acid binding protein 7(FABP7),analyze it with bioinformatics,and detect the mRNA expression level of FABP7 in cattle tissues.The CDS sequence of FABP7 gene in Red Steppe cattle was cloned by RT-PCR,and a variety of biological software and online tools were used for bioinformatics analysis.The mRNA expression level of FABP7 gene in various tissues was detected by qPCR technology.In this study,the CDS of FABP7 gene was 399 bp in length encoding 132 amino acids with a protein molecular weight of 14.96 ku and a theoretical isoelectric point of 5.38,which was a hydrophilic protein.By NCBI-BLAST comparison,the nucleotide sequence homology of the FABP7 gene of Red Steppe cattle shared 99%,98%,94%,92%,85%and 85%identity with Bos taurus,Ovis aries,Sus scrofa,Homo sapiens,mouse,and Gallus.The phylogenetic tree found that the Red Steppe cattle had the closest relationship with Bos taurus and Ovis aries,and the farthest relationship with Gallus.The protein sequence had 1 disulfide bonds,14 phosphorylation sites,1 N-glycosylation sites,no signal peptide and transmembrane region.In the secondary structure and tertiary structure of the protein,it was found that the FABP7 protein mainly has twoα-helix structures and tenβ-turns,which was a mixed protein.The FABP7 gene had the highest expression in small intestine and was moderately expressed in the heart,fat and stomach.
作者
吕阳
曹阳
高一
刘理想
薛佳佳
张国梁
LV Yang;CAO Yang;GAO YI;LIU Lixiang;XUE Jiajia;ZHANG Guoliang(Branch of Animal Husbandry,Jilin Academy of Agricultural Sciences,Jilin Gongzhuling 136100,China)
出处
《中国畜牧杂志》
CAS
北大核心
2019年第9期48-52,57,共6页
Chinese Journal of Animal Science
基金
国家肉牛牦牛产业技术体系公主岭综合试验站（CARS-37）
国家重点研发计划项目（2018YFD0501802）
吉林省农业科技创新工程研究生基金项目
肉用草原红牛培育：畜禽品种培育项目
