摘要
为了获得巴马香猪脂肪酸结合蛋白4(FABP4)基因的编码区序列(CDS),并探究FABP4基因在巴马香猪不同器官/组织中的mRNA表达水平,试验以巴马香猪背最长肌为材料,提取总RNA并扩增出FABP4基因的CDS,运用在线分析系统进行生物信息学分析,并利用实时荧光定量PCR技术检测巴马香猪心脏、脾脏、肝脏、肾脏、肺脏、背最长肌和胰腺等器官/组织中FABP4基因的mRNA表达水平。结果表明:成功克隆得到巴马香猪FABP4基因的CDS,长度为488bp,与GenBank中公布的猪FABP4基因同源性为99.6%,并且存在2个突变位点,第421位点处G→A为同义突变,第462位点处A→G导致赖氨酸突变为精氨酸;巴马香猪FABP4基因CDS与猪、人、黑猩猩、猕猴、牛和家鼠的同源性分别为99.6%、89.3%、89.3%、88.7%、87.3%和82.8%,且与普通猪的遗传距离最近,聚类为一支;巴马香猪FABP4蛋白氨基酸组成中苏氨酸和缬氨酸含量较高,均占氨基酸总数的10.6%;FABP4蛋白为亲水蛋白,包含一个Lipocalin超家族结构域;巴马香猪FABP4蛋白的二、三级结构包含α-螺旋、延伸链、β-折叠和无规则卷曲;另外,巴马香猪背最长肌FABP4基因的mRNA表达量显著高于其他器官/组织(P<0.05),肾脏中的表达量也较高,胰腺中的表达量最低。说明FABP4基因可作为探究巴马香猪脂肪沉积的主要候选基因之一。
In order to obtain the coding sequence(CDS)of fatty acid binding protein 4(FABP4)gene in Bama Xiang pig,and to explore the mRNA expression level of FABP4 gene in different tissues of Bama Xiang pig,the total RNA was extracted from the longissimus dorsi muscle and the FABP4 gene CDS was amplified.Bioinformatics analysis was carried out by online analysis system,and the mRNA expression levels of FABP4 gene in Bama Xiang pig heart,spleen,liver,kidneys,lungs,longissimus dorsi muscle and pancreas were detected by fluorescence quantitative PCR.The results showed that the CDS of FABP4 gene of Bama Xiang pig successfully cloned with a length of 488 bp,and its homology was 99.6%with the pig gene was published by GenBank.There were two different sites,the G→A was a missense mutation in the 421 th position,the A→G also was a missense mutation in the 462 th position,which caused the alanine turns to leucine.The homology of FABP4 gene in Bama Xiang pig was 99.6%,89.3%,89.3%,88.7%,87.3%and 82.8%with pigs,humans,chimpanzees,macaques,cattle and mice.It had the closest genetic distance to the common pig and clusters into one branch.The contents of threonine and valine in the amino acid composition of FABP4 were higher,accounting for 10.6%of the total amino acids.FABP4 was a hydrophilic protein,and contained a lipocalin superfamily domain.The secondary and tertiary structures of FABP4 protein includedα-helix,elongation chain,β-fold and irregular curl.In addition,the mRNA expression of FABP4 gene in Bama Xiang pig’s longissimus dorsi muscle was significantly higher than that in other tissues,the expression level in kidney was also higher,and the expression level in pancreas was the lowest.It indicated that the FABP4 gene could be used as one of the main candidate genes to explore fat deposition in Bama Xiang pig.
作者
谢红月
潘鹏
罗云彦
胡旭旭
甄锐
蒋钦杨
黄艳娜
XIE Hongyue;PAN Peng;LUO Yunyan;HU Xuxu;ZHEN Rui;JIANG Qinyang;HUANG Yanna(College of Animal Science and Technology,Guangxi University,Nanning 530004,China)
出处
《黑龙江畜牧兽医》
CAS
北大核心
2020年第21期19-24,共6页
Heilongjiang Animal Science And veterinary Medicine
基金
国家自然科学基金项目(31760672,31460606)
作者简介
谢红月(1995—),女,硕士研究生,研究方向为动物营养与肉质调控,2996970767@qq.com;通信作者:黄艳娜(1980—),女,副教授,博士,硕士生导师,研究方向为动物营养与肉质调控,huangyn@gxu.edu.cn.
