摘要： 为了丰富牦牛FAM134B 基因研究的基础数据,进一步探讨FAM134B 基因的生理功能,通过PCR扩增和测 序技术并结合生物信息学分析软件,对牦牛FAM134B 基因进行克隆、测序以及相关生物信息学分析.克隆获得1 079 bp 的牦牛FAM134B 基因,其中CDS区全长1 071 bp(GenBank 登录号:KM587693),编码356个氨基酸残基组成的蛋白质.与普通牛比对,牦牛FAM134B 基因在CDS区存在3个碱基突变,其中第727位上碱基C→T的突变导致密码子CCC→TCC,使编码的氨基酸由脯氨酸变成丝氨酸.牦牛FAM134B 基因编码蛋白的分子式为C₁₇₀₅H₂₆₈₆N₄₄₈O₅₇₅S₁₃ ,分子量为39.077 5 kDa,理论等电点(pI)为4.46,消光系数为24 450.不稳定系数为46.85,疏水指数为79.47,平均亲水性为-0.439,属不稳定可溶性酸性蛋白质.二级结构以无规卷曲和α-螺旋为主,其中α-螺旋占23.88%,无规卷曲占74.72%,属混合类蛋白质.亚细胞定位结果显示:FAM134B 编码的蛋白在内质网、细胞质膜、空泡、细胞核、高尔基体、细胞骨架和线粒体中分别占30.4%,21.7%,17.4%,17.4%,4.3%,4.3%,4.3%,推测其可能在物质的运输以及辅因子的生物合成等过程中发挥离子通道载体以及信号转导和转录调控的作用.系统发育树分析表明,牦牛FAM134B氨基酸序列与普通牛、绵羊、小鼠、褐家鼠、猕猴、黑猩猩、人、非洲爪蟾的FAM134B氨基酸序列的同源性分别为99.7%,97.2%,87.1%,86.8%,90.4%,90.2%,90.4%,57.9%,物种间的同源性较高,其系统进化的情况与亲缘关系的远近相一致,说明FAM134B 基因的编码区在进化的过程中比较保守.FAM134B 基因的成功克隆和分析为揭示牦牛FAM134B 基因的遗传特性提供了理论依据.

关键词: 牦牛, FAM134B 基因, CDS区克隆, 生物信息学分析

Abstract: In order to enrich basic data in yak FAM134B gene and further explore the physiological function of FAM134B gene.By use of the PCR amplification and sequencing and bioinformatics analysis software to complete the yak FAM134B gene cloning,sequencing and bioinformatics analysis.The 1 079 bp length FAM134B gene in yak was got by cloning,including the 1 071 bp length CDS region(GenBank accession No.:KM587693),and encoding 356 amino acids.Alignment with CDS and amino acid sequence of cattle FAM134B gene,three base mutations were found.The 727^th base C was mutated to T and caused codon CCC variable TCC encoding amino acid from proline to serine.The formula of protein encoded by FAM134B gene in yak was C₁₇₀₅H₂₆₈₆N₄₄₈O₅₇₅S₁₃,and the molecular weight was 39.077 5 kDa,the theory isoelectric point was 4.46,the extinction coefficient was 24 450,the instability index was 46.85,the aliphatic index was 79.47,and the grand average of hydropathicity was-0.439.It was an unstable and soluble protein.The secondary structure of FAM134B was mainly α-helices and random coil,α-helices was 23.88% and random coil was 74.72%,FAM134B was belongs to mixed type of protein.Subcellular localization of FAM134B was in the endoplasmic reticulum(30.4%), plasma membrane(21.7%),vacuolar(17.4%),nuclear(17.4%),golgi(4.3%),cytoskeletal(4.3%),and mitochondrial(4.3%),speculated that it might play ion channel carrier and signal transduction and transcriptional regulation in material transport and cofactor biosynthesis process.The amino acid homology of FAM134B compared Yak to Bos taurus,Ovis aries,Mus musculus,Rattus norvegicus,Macaca mulatta,Pan troglodytes,Homo and Xenopus laevis sapiens were 99.7%,97.2%,87.1%,86.8%,90.4%,90.2%,90.4%,57.9%.There was a high homology among different species,and phyletic evolution were same as their genetic relationship.The research indicated that the FAM134B gene coding region was conservative in the course of evolution.The gene was cloned and analyzed,which provided the theoretical basis for revealing the genetic characteristics of yak FAM134B gene.

Key words: Yak, FAM134B gene, CDS cloning, Bioinformatics

中图分类号: 

• Q78

• S823