猪EIF2S3基因克隆测序及其组织表达谱分析  

Cloning,sequencing of porcine gene EIF2S3 and its tissue expression profile

作　　者：龙熙[1] 张廷焕 赵久刚[1] 蓝静[1] 柴捷[1] 郭宗义[1] 王金勇[1] 张亮[1] LONG Xi;ZHANG Ting-huan;ZHAO Jiu-gang;LAN Jing;CHAI Jie;GUO Zong-yi;WANG Jin-yong;ZHANG Liang(Chongqing Academy of Animal Science,Chongqing 402460,China)

机构地区：[1]重庆市畜牧科学院,重庆402460

出　　处：《南方农业学报》2018年第10期2062-2069,共8页Journal of Southern Agriculture

基　　金：国家自然科学基金面上项目(31771376);重庆市基本科研业务费专项项目(16435);重庆市科研院所绩效激励引导专项项目(cstc2017jxjl0065)

摘　　要：【目的】克隆猪EIF2S3基因mRNA全长,并进行生物信息学分析及检测其在猪各组织中的表达特征,为研究真核翻译起始因子2(eIF2)蛋白γ亚基的生物学功能打下基础。【方法】以荣昌猪胸腺mRNA为模板,采用RCAE-PCR克隆EIF2S3基因的m RNA全长序列,在线完成各种生物信息学分析后,利用实时荧光定量PCR分析猪EIF2S3基因的组织表达特征。【结果】猪EIF2S3基因mRNA全长1813 bp,其中蛋白质编码区(CDS)长1419 bp,5'UTR区、3'UTR区分别长14和380 bp,编码472个氨基酸。EIF2S3蛋白分子量51.1 kD,理论等电点(pI)8.62,包含58个带正电的氨基酸和52个带负电的氨基酸,其前体蛋白靠近N端区域有一个强疏水区;EIF2S3蛋白的空间结构由α-螺旋、β-折叠、延伸及无规则卷曲构成。EIF2S3蛋白氨基酸序列与狗和大白鼠的相似性最高(99.6%),其次是马、猫、牛、羊和人类(99.4%),与斑马鱼的相似性最低(96.6%)。EIF2S3基因在猪的心脏、胸腺和淋巴组织中表达量较高,在脾脏和肌肉中呈中度表达,在肾脏中的表达量较低,而在肝脏中基本不表达。【结论】猪EIF2S3基因全长1813 bp,在核酸和氨基酸水平上与其他物种的EIF2S3基因高度同源,尤其与狗和大白鼠的遗传距离最近;其在猪心脏、胸腺和淋巴组织中的表达量较高,在肝脏中基本不表达,据此推测EIF2S3参与合成的蛋白更多地参与心脏泵血和免疫相关功能,基本不发挥能量代谢功能。【Objective】The aim of the present study was to clone the full-length of gene EIF2S3 mRNA in pig,conduct bioinformatics analysis and detect its expression characters in various tissues and lay the foundation for studying the biological function of theγsubunit of eukaryotic translation initiation factor 2(eIF2)protein.【Method】Thymus mRNA of Rongchang pig was as template,RCAE(Rapid-amplification of cDNA ends)-PCR was used to clone full-length sequence of gene EIF2S3 mRNA.After various bioinformatics analysis was conducted online,real-time fluorescence quantitative PCR was applied to study the expressions of EIF2S3 in different tissues.【Result】The results showed that the EIF2S3 gene mRNA of pig was 1813 bp in length.This sequence consisted of 1419 bp of CDS,14 bp of 5'UTR and 380 bp of 3'UTR.A total of 472 amino acids were encoded.Molecular weight of EIF2S3 protein was 51.1 kD,and theoretical isoelectric point(pI)8.62.The EIF2S3 protein contained 58 positively charged amino acids and 52 negatively charged amino acids.A strong hydrophobic region near the N-terminal region of EIF2S3 precursor protein was found.The tertiary structure of EIF2S3 comprisedα-helices,β-folds,extensions and random coils.Amino acid sequence of EIF2S3 protein had the highest similarity with Canis lupus familiaris and Rattus norvegicus(99.6%),followed with Equus caballus,Felis catus,Bos Taurus,Ovis aries and Homo sapiens(99.4%),and it had the lowest similarity with Danio rerio(96.6%).The high expression levels of EIF2S3 were in the heart,thymus and lymphonodi.Medium expression levels were observed in spleen and muscle.The expression of EIF2S3 in kidney was low and it basically not expressed in the liver.【Conclusion】The EIF2S3 gene of pig is 1813 bp in length.It is highly homology with other species in nucleotide and amino acid level and has the closest genetic relationship with C.lupus familiaris and R.norvegicus.EIF2S3 has high expression levels in the heart,thymus and lymphonodi,but nearly no expression in liver.Thus,it is s

关 键 词：猪 真核翻译起始因子2(eIF2) EIF2S3基因 克隆 序列分析 组织表达谱 

分 类 号：S828.89[农业科学—畜牧学]