刀鲚MyoG基因克隆和组织表达分析

Cloning and Tissue Expression Analysis of MyoG Gene in Coilia nasus

为了探究刀鲚(Coilia nasus)MyoG基因的生物学功能及组织表达特征,采用RACE克隆其MyoG基因cDNA全长序列,并进行生物信息学分析,从GenBank中选择硬骨鱼类、两栖动物、鸟类和哺乳动物4种物种的MyoG氨基酸序列,通过DNAMAN进行氨基酸序列比对,以MEGA 7.0中的邻接法(NJ)构建系统发育进化树,应用实时荧光定量PCR检测分析刀鲚MyoG基因在不同组织的表达情况。结果表明,刀鲚MyoG基因cDNA全长1836 bp,包括118 bp的5'非编码区(5'-UTR)、944 bp的3'非编码区(3'-UTR)和774 bp的开放阅读框(ORF),共编码257个氨基酸残基。刀鲚MyoG氨基酸序列具有MRFs典型的碱性螺旋-环-螺旋(bHLH)结构域,与其他鱼类的MyoG氨基酸序列同源性较高,其中与大西洋鲱(Monopterus albus)的MyoG氨基酸序列相似性最高。刀鲚MyoG基因在其肌肉组织中的相对表达量最高,在心脏和脾脏中最低。综上,本研究成功克隆了刀鲚MyoG基因cDNA全长序列,并对其生物信息学及组织表达特征进行了分析,研究结果为进一步探究刀鲚MyoG基因功能特征提供了参考。

To explore the biological functions and tissue expression characteristics of MyoG in Coilia nasus,this study used RACE to clone the full-length cDNA sequence and conducted bioinformatics analysis.The MyoG amino acid sequences of four species,including bony fish,amphibians,birds,and mammals,were selected from GenBank.The amino acid sequences were aligned using DNAMAN,and a phylogenetic tree was constructed using the Neighbor Joining(NJ)in MEGA 7.0.The expression of MyoG gene in different tissues of C.nasus was detected and analyzed using Quantitative Real-time PCR.The results showed that the full-length cDNA of MyoG gene in the C.nasus was 1836 bp,including 118 bp of 5'-UTR,944 bp of 3'-UTR,and 774 bp of open reading frame(ORF),encoding a total of 257 amino acid.The MyoG amino acid sequence of the C.nasus had the typical basic helix loop helix(bHLH)domain of MRFs,and had high homology with the MyoG amino acid sequences of other fish species,with the closest relationship being with the Monopterus albus.The relative expression level of MyoG gene in the muscle tissue of the C.nasus was the highest,while it was lower in the heart and spleen.In conclusion,this study successfully cloned the full-length cDNA sequence of MyoG gene in the C.nasus,and analyzed the bioinformatics and tissue expression characteristics of MyoG gene.The research results provides a reference for further exploring the functional characteristics of MyoG gene in C.nasus.