甘肃马鹿肌细胞生成素(MyoG)基因启动子区序列克隆与分析

Cloning and Analysis of the Promoter Region Sequence of MyoG Gene in Gansu Wapiti

MyoG基因在肌肉发生过程中具有中心调节作用。根据GeneBank中已发表的猪、人、小鼠和牛的MyoG基因5'侧翼和部分第1外显子序列设计PCR引物,采用Touch-Down PCR技术扩增了甘肃马鹿MyoG基因的启动子区序列,构建了重组克隆载体pMD18-T-MyoG,并通过PCR扩增、限制性酶切对阳性克隆进行了鉴定、测序及生物信息学分析。结果表明,甘肃马鹿MyoG基因启动子区序列长685bp,其与猪、人和小鼠的同源序列相似性分别为88.1%、84.9%和82.6%,且不同物种间保守性较强。本研究为进一步探讨甘肃马鹿MyoG基因的表达与调控机制奠定了基础。

MyoG gene was reported to play a core role in regulating myogenesis. According to 5＇flank regions and partial sequences of the MyoG gene in Sus scrofa, Bos taurus, Homo sapiens, and Mus musculus published in GenBank, the homogeneous primers were designed to amplify the promoter region of Gansu wapiti MyoG by Touch-Down PCR. The PCR product was cloned into pMD 18-T vector, and then was transformed into E.coli DH5 α. Positive clones were identified by using PCR, restriction endonuclease, sequencing and bioinformatics analysis. The results showed that the length of promoter region sequence in Gansu wapiti MyoG was 685bp, and the similarity was 88.1%, 84.9% and 82.6% with those of Sus scrofa, Homo sapiens and Mus musculus respectively. It indicates that the promoter region sequence of MyoG was comparatively conservative among different species. This work would lay the foundation for further study on regulating and expressing mechanism of MyoG in Gansu wapiti.