绵羊c-Myc的克隆与序列分析

Cloning and Coding Sequence Analysis of Sheep c-Myc Gene

参照GenBank上绵羊c-Myc序列设计引物,利用RT-PCR技术从60d左右胎羊生殖嵴总RNA中扩增得到绵羊c-Myc基因编码区序列,其为包括终止密码子在内的1320 bp,编码有439个氨基酸。同源性分析结果显示,绵羊c-Myc基因编码区与牛、猪、猫、人、大鼠、小鼠、鸡、爪蟾和斑马鱼的核苷酸序列同源性分别为98.0%、94.1%、92.4%、90.4%、86.9%、86.2%、75.2%、66.4%和63.9%,其氨基酸序列同源性分别为98.0%、94.0%、92.5%、90.3%、87.2%、86.5%、72.2%、65.5%和64.6%;生物信息学软件分析显示,绵羊c-Myc含有细胞核定位模体和HLH结构域。本研究为进一步研究c-Myc基因的功能及探讨其中绵羊体细胞重编程中的作用奠定了基础。

The c-Myc gene was amplified from E60 sheep genital ridge mRNA, and ligated with linear vector pMD-18T. The sequencing results of sheep c-Mye gene （already been logged in GenBank, indicated that it＇s CDS, including stop coden, was in a length of 1320bp, encoding 439 amino acids, and contained nuclear localization motifs and HLH domain showed by bio-software analysisng. Sheep c-Myc gene also shared 98.0% ,94.1% ,92.4% ,90.4% ,86.9% ,86.2% ,75.2% ,66.4% and 63.9% nucleotide homology and 98.0%, 94.0% ,92.5% ,90.3% ,87.2% ,86.5% ,72.2% ,65.5% and 64.6% amino acid identity with Bos taurus, Sus scrofa, Felis catus, Homo sapiens, Rattus norvegicus, Mus musculus, Gallus gallus, Xenopus laevis and Danio rerio respectively. The successful cloning of e-Mye shows that the gene in the structure and function is very＂ conservative and helpful to study gene function and role in somatic reprogramming in future.