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陆川猪MyoD1基因克隆及组织表达分析OA北大核心CSTPCD

Cloning and Tissue Expression Analysis of MyoD1 Gene in Luchuan Pig

中文摘要英文摘要

为探究陆川猪骨骼肌生长发育过程中MyoD1 基因所起作用,对陆川猪成肌细胞决定基因1(MyoD1)进行克隆,展开生物信息学分析,对其在陆川猪不同组织中的表达进行分析,以NCBI上已公布的野猪MyoD1 基因序列为模板,进行特异性引物设计,克隆陆川猪MyoD1 基因CDS区,并与NCBI已公布的野猪、牛、绵羊、人、小鼠、大鼠基因序列进行相似性比对,由此构建系统进化树,通过在线软件开展生物信息学分析,而后使用实时荧光定量PCR检测在陆川猪不同组织中MyoD1 基因的相对表达量.陆川猪MyoD1 基因CDS区全长960 bp,共编码319 个氨基酸,碱基突变共存在5 处,与野猪、牛、绵羊、人类、小鼠、大鼠的 MyoD1 基因 CDS 序列相似性分别为 99.2%,93.0%,92.3%,90.0%,84.3%,84.0%,其中与野猪相似性最高,表明二者亲缘关系最为接近;陆川猪MyoD1 基因原子总数为4680,蛋白的分子质量为33.99 ku,分子式为C1457 H2296 N442 O471 S14;不稳定系数为63.87,表明其缺乏稳定性;等电点为5.63,属于酸性蛋白;不存在跨膜结构,存在 35 处磷酸化位点及 1 处糖基化位点;蛋白二级结构以无规则卷曲为主,占比60.69%.陆川猪MyoD1 基因在背最长肌中的表达量最高,且显著高于其他组织,表达量最低的组织为肾脏.陆川猪各组织中均有MyoD1 基因表达,且主要在背最长肌中表达,由此推测,MyoD1 基因在陆川猪肌肉生长发育过程中可能起到至关重要的作用.

Luchuan pig myoblast determining gene 1(MyoD1)was cloned,bioinformatics analysis was performed,and its expression was analyzed in different tissues of Luchuan pigs,to explore the role of MyoD1 gene in the growth and development of skeletal muscle.Published MyoD1 gene of wild boar sequence on NCBI as the template,designed specific primer,cloned the CDS region of MyoD1,and compared the similarity with wild boar,cattle,sheep,human,mouse and rat gene sequence,constructed system tree for bioinformatics analysis by online software.Then,the relative expression of MyoD1 gene in different tissues of Luchuan pigs by RT-PCR.The CDS of MyoD1 gene in Luchuan pig was 960 bp,encoding 319 amino acids,five base mutations.The similarities of MyoD1 genes between Luchuan pig and wild boar,cattle,sheep,human,mouse and rats were 99.2%,93.0%,92.3%,90.0%,84.3%and 84.0%,respec-tively.Total number of MyoD1 gene atoms in Luchuan pigs was 4680,molecular mass of the protein was 33.99 ku,molecular formula was C1457 H2296 N442 O471 S14;the instability coefficient was 63.87,indicating a lack of stability;isoelec-tric point was 5.63,which belongs to the acidic protein;no transmembrane structure existed,found 35 phosphorylation sites and 1 glycosylation site;protein secondary structure was dominated by an irregular coil,which accounting for 60.69%.The MyoD1 gene in Luchuan pigs had the highest expression in the longest dorsal muscle,which was sig-nificantly higher than in other tissues,while kidney had the lowest expression.MyoD1 gene was expressed in all tis-sues of Luchuan pigs and mainly in the longest dorsal muscle,we speculated that MyoD1 gene might play a crucial role in the muscle growth and development of Luchuan pigs.

高泽仁;潘鹏丞;徐文文;陈宝剑;刘明君;关志惠;谢炳坤;覃兆鲜

广西大学 动物科学技术学院,广西 南宁 530004广西农业职业技术大学 畜牧研究院,广西 南宁 530001广西大学 动物科学技术学院,广西 南宁 530004||广西农业职业技术大学 畜牧研究院,广西 南宁 530001

畜牧业

陆川猪MyoD1基因生物信息学分析基因克隆组织表达

Luchuan pigMyoD1 geneBioinformatics analysisGene cloningOrganizational expression

《华北农学报》 2024 (002)

168-173 / 6

广西重点研发计划(桂科AB21196060);中央引导地方科技发展专项(桂科ZY21195052);广西农业科技自筹经费项目(Z2022119);国家现代农业产业技术体系广西创新团队建设专项资金(nycytxgxcxtd-2023-15-01)

10.7668/hbnxb.20194657

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