为了克隆出山羊c-Myc基因的CDS区序列,本研究利用RTPCR技术,参照GenBank报道的绵羊、牛、猪、马等的c-Myc基因CDS区序列设计出特异性引物,从山羊胚胎皮肤组织中扩增出山羊c-Myc CDS区序列并进行序列分析。结果表明,山羊c-Myc基因CDS区全长1320bp(包括终止密码子),与绵羊c-Myc基因核苷酸序列(GenBank登录号:Z68501.1)比对仅有10处存在差异,其核苷酸序列同源性为99.17%,推导的氨基酸序列同源性为99.09%;与其他物种进行同源性分析结果显示,山羊c-Myc基因CDS区与猪、狼、人、大鼠、小鼠的核苷酸序列同源性分别为91.2%、92.0%、90.2%、86.9%、86.0%,推导的氨基酸序列同源性分别为93.6%、96.1%、92.3%、91.1%、89.6%;生物信息学软件分析结果表明,山羊c—Myc蛋白产物定位于细胞核并含有HLH结构域。本研究为进一步了解c-Myc基因的功能及探讨其在山羊体细胞重编程过程中的作用奠定了基础。
In order to clone sequence of the coding region of the goat c-Myc gene, this study designed specific primers by refeting to c-Myc genes coding sequence reported in GenBank including sheep,cattle, pigs, horses, etc. , and then amplified c-Myc coding sequence from embryonic skin tissue of goats by using RT-PCR technique and had sequence analysis. The result indicated that CDS of goat c-Myc gene had 1320 bp length including the stop codon. For only 10 differences comparing to sheep c-Myc gene nucleotide sequence (GenBank accession number: Z68501.1), homology reached 99.17 % and homology of deduced amino acid sequence reached 99.09%. Analysis of homology with other species showed that homology of goat c-Myc gene coding region nucleotide sequence was 91.2 %, 92.0%,90.2 %, 86.9 %,86.0 % with pigs, wolves, humans, rats, mice, and homology of deduced amino acid sequence was 93.6 %, 96.1 %, 92.3%,91.1%, 89.6 %. Using bioinformatics software analysis showed that goat c-Myc protein was localized in the nucleus and contained a HLH domain. This study further researched the function of c-Myc gene and laid the foundation of investigating the effect in the process of goat somatic cell reprogramming.