豪猪MITF-M转录本测序及其生物信息学分析  

Transcript sequencing and bioinformatics analysis of MITF-M gene in porcupines

作　　者：唐克桐 秦廷洋 龙红 张仕斌 TANG Ketong;QIN Tingyang;LONG Hong;ZHANG Shibin(Laboratory Animal Center,Zunyi Medical University,Zunyi 563003,China)

机构地区：[1]遵义医科大学实验动物中心,贵州遵义563003

出　　处：《黑龙江畜牧兽医》2022年第10期128-132,138,共6页Heilongjiang Animal Science And veterinary Medicine

基　　金：遵义市科技局遵义医科大学科学技术联合资金项目(遵市科合HZ字[2020]57号)。

摘　　要：为获得豪猪MITF-M转录本序列,并对其进行生物信息学分析,试验首先将人工饲养豪猪的耳廓皮肤组织样本提取总RNA后进行无参转录组测序,对获得的所有MITF转录本通过序列比对分析筛选豪猪MITF-M转录本序列,然后采用生物信息学在线软件对豪猪MITF-M基因编码蛋白的理化性质、氨基酸组成、亚细胞定位、二级结构、三级结构、信号肽和跨膜区域进行预测和分析,最后分析豪猪与大鼠、鸡、绵羊等9个物种MITF-M蛋白的同源性,并采用邻接法构建MITF-M蛋白的系统进化树。结果表明:获得豪猪MITF-M转录本序列(已上传至GenBank,登录号为MW84037),序列全长为4772 bp,含9个外显子,CDS长度为1260 bp(229~1488 bp),编码419个氨基酸。豪猪MITF-M蛋白化学分子式为C_(2010)H_(3263)N_(585)O_(652)S_(26),共由6536个原子组成;相对分子质量为46890.11;理论等电点为5.82,为酸性蛋白质;脂肪氨基酸系数为79.64,不稳定指数为69.40,亲水指数为-0.617。豪猪MITF-M蛋白共由20种氨基酸构成,其中亮氨酸含量最高(占比为11.5%),而色氨酸含量最低(占比为0.2%);带正电荷(天冬氨酸、谷氨酸)和负电荷(精氨酸、赖氨酸)的氨基酸残基数目分别为52个和44个;二级结构由α-螺旋、延伸链、β-转角和无规则卷曲构成,其中α-螺旋占比为37.47%,延伸链占比为7.40%,β-转角占比为2.86%,无规则卷曲占比为52.27%;三级结构中存在左右对称的CLEAR-box结构;豪猪MITF-M蛋白不具有信号肽,不存在跨膜区域;豪猪MITF-M蛋白的氨基酸序列与人MITF-M蛋白相似性最高,为95.7%;与猪MITF-M蛋白变异性最低,仅为4.2%。豪猪与同属啮齿目的大鼠、小鼠的进化距离较近,与物种分类相吻合。说明成功获得了豪猪MITF-M转录本序列,其编码蛋白符合转录因子在细胞核中发挥作用的结构特点。In order to obtain the porcupine MITF-M transcription sequence and conduct bioinformatics analysis,in this experiment,total RNA was extracted from the auricular-skin tissue samples of artificially reared porcupine and transcriptome sequencing performed without reference.All the obtained MITF gene transcripts were screened by sequence alignment analysis for porcupine MITF-M transcripts.Then,the physicochemical properties,amino acid composition,subcellular localization,secondary structure,tertiary structure,signal peptide and transmembrane region of the protein encoded by the porcupine MITF-M gene were predicted and analyzed by bioinformatics online software.Finally,the homology of MITF-M protein between porcupine and 9 species such as Rattus norvegicus,Gallus gallus and Ovis aries was analyzed.Neighbor-joining(NJ)method was used to construct the phylogenetic tree of MITF-M protein.The results showed that the transcription sequence of MITF-M of porcupine(uploaded to GenBank,entry number is MW84037)was 4772 bp,containing 9 exons,and CDS length was 1260 bp(229-1488 bp),encoding 419 amino acids.The molecular formula of porcupine MITF-M protein was C_(2010)H_(3263)N_(585)O_(652)S_(26),consisting of 6536 atoms.The relative molecular weight was 46890.11.The theoretical isoelectric point was 5.82,which was acidic protein.The amino acid coefficient of fat was 79.64,the instability index was 69.40,and the hydrophilic index was-0.617.Porcupine MITF-M protein was composed of 20 amino acids,among which leucine was the highest,accounting for 11.5%,while tryptophan was the lowest,accounting for only 0.2%.The number of amino acid residues with positive(aspartic acid,glutamic acid)and negative(arginine,lysine)charges was 52 and 44,respectively.The secondary structure was composed ofα-helix,extension chain,β-rotation and random curl,of which theα-helix accounted for 37.47%;the extension chain accounted for 7.40%;theβ-rotation accounted for 2.86%,and the random curl accounted for 52.27%.There was a leftright symmetric CLEAR-box

关 键 词：豪猪 MITF-M基因 转录本 生物信息学分析 基因注释 

分 类 号：Q71[生物学—分子生物学]