赤眼鳟MyoG基因的克隆与组织表达分析  

Molecular cloning and tissue expression analysis of MyoG gene in Squaliobarbus crriculus

作　　者：张燕萍[1] 邵芳[1] 周峰[1] 徐建荣[1] 顾志良[1] 

机构地区：[1]常熟理工学院生物与食品工程学院,江苏常熟215500

出　　处：《南京农业大学学报》2014年第5期117-122,共6页Journal of Nanjing Agricultural University

基　　金：教育部留学回国人员科研启动基金(第42批);苏州市科技基础设施建设项目(SZSD201001)

摘　　要：本研究以赤眼鳟肌肉总RNA为模板,采用RT-PCR、3'-RACE和5'-RACE的方法,获得了赤眼鳟MyoG基因的全长cDNA序列。该基因含有完整的开放阅读框,编码274个氨基酸;编码的蛋白含有典型的bHLH结构。序列分析表明:赤眼鳟MyoG基因与其他鱼类核苷酸和氨基酸同源性都比较低,其中最高的是鲤鱼,核苷酸同源性和氨基酸同源性分别为78.42%和74.09%。进化系统分析表明,赤眼鳟MyoG基因在进化上高度保守。采用半定量RT-PCR方法,检测赤眼鳟MyoG mRNA在肌肉、脑、肠和肝脏等不同组织的表达情况。结果表明:MyoG mRNA在肌肉中的表达量最高,肾、心和脑中的表达量次之,鳃中有微量表达,脾脏中有痕量表达,在肝脏和肠中不表达。揭示MyoG除了在肌肉中起作用外,在其他组织中也可能参与相关的作用。In the current study, Squaliobarbus crriculus MyoG gene fragments were amplified by RT-PCR, 3＇ RACE and 5＇ RACE methods. A full-length of 1 628 bp MyoG cDNA sequence in S. crriculus was assembled,which includes a complete ORF encoding a 274 amino acids peptide with a typical bHLH structure in the protein. Sequence analysis indicated that the S. crriculus MyoG gene exhibited low similarity of nucleotide and amino acid with other fishes, and the highest one of nucleotide similarity and amino acid similarity is Cyprinus carpio,which was 78.42% and 74.09% ,respectively. A phylogenetie tree of 13 species was constructed using the Neighbor-Joining method by MAGE 4.2, and the phylogenetic relationship of the 13 species was revealed. It indicated that the S. crriculus MyoG gene was highly conserved in evolution. MyoG gene was detected in all eight examined tissues by semi-quantitative RT-PCR,MyoG mRNA expressed highest in muscle, followed in kidney, heart and brain, weakly expressed in gill, and traced in spleen,implying that the fish＇ MyoG gene would play an important role in muscle development.

关 键 词：赤眼鳟 MYOG基因 RACE 基因克隆 组织表达 

分 类 号：Q38[生物学—遗传学] S917.4[农业科学—水产科学]