摘要
根据GenBank中公布的绵羊、水牛、猪、小鼠及人的肌细胞生成素基因(myogenin)DNA序列,设计了3对引物,采用PCR扩增和克隆技术,首次从甘肃马鹿血液基因组中获得了myogenin基因序列,并利用生物信息学方法对甘肃马鹿该基因核苷酸及其编码氨基酸序列的结构特征进行初步预测和分析,进一步基于该基因氨基酸序列构建了15个物种的分子系统进化树。结果表明,获得的甘肃马鹿myogenin基因序列大小为3168bp(GenBank登录号:FJ746497),包括3个外显子、2个内含子、部分5'UTR和3'UTR,其序列包括1个684bp的开放阅读框,编码227个氨基酸,myogenin蛋白理论分子质量为25.2493kDa,PI为5.68,不稳定系数是66.22,属于酸性不稳定蛋白质。进一步分析发现,该序列不含有信号肽和跨膜区域,含有11个广泛磷酸化位点,第1～136个氨基酸为甘肃马鹿myogenin基因bHLH保守结构功能域,二级结构以无规则卷曲和α-螺旋为主。序列相似性及分子系统进化分析显示,甘肃马鹿myogenin基因与反刍动物山羊、绵羊和水牛的亲缘关系最近。甘肃马鹿myogenin基因的克隆及序列结构特征的生物信息学预测为进一步探讨鹿科动物肌肉生长发育的分子调控机理奠定了理论基础。
In the current study, three primers were designed according to the result of comparation among myogenin gene in Ovis aries （DQ453548）, Bubalus bubalis （EF636460）, Sus scrofa （SSU14331）, Mus musculus （M95800） and Homo sapiens （X62155） published in GenBank. The partial DNA sequence （3168 bp） were obtained for myogenin gene by PCR and cloning technique from the blood total DNA of wapiti Cervus elaphus kansuensis, then the structural characteristics of nueleotide and amino acid sequence were initially predicted and analyzed by the method of bioinformatics. Phylogenetic tree of myogenin gene among 15 species based on the amino acid sequence was constructed. The results showed that the sequence of wapiti myogenin gene consisted of 3 exons, 2 introns and partial of 5＇UTR （381 bp） and 3＇UTR （696 bp） untranslated region, with an open reading frame of 684 bp, which encodes 227 amino acids. The estimated molecular weight of myogenin protein is 25. 2493 kDa, with an isoeleetric point of 5.68 and 62.22 in instability index, belonging to the unstable acid pro- tein. Further structure prediction indicated that the myogenin protein had no signal peptide and transmembrane domain, which includes 11 generic phosphorylation sites in wapiti myogeuin protein, and a bHLH conserved functional domain was located at No. 1 -136 amino acids, as well as the main component of all secondary structures includes or-helix and random coil. The similarity comparison and phylogenetic tree indicated that the evolution distance of wapiti myogenin was the most homogeneous to goat, sheep and cattle belonging to ruminant. This study will pave the way for functional characterization of the myogenin gene in wapiti. All researches made an essential foundation for myogenin expression, evolution and polymorphism analysis for the future.
出处
《四川动物》
CSCD
北大核心
2013年第1期51-59,共9页
Sichuan Journal of Zoology
基金
国家科技基础条件平台建设项目(2005DKA21102):特种经济动物标准化整理
整合及共享试点
国家科技支撑计划项目(2006BAD06B06)
关键词
甘肃马鹿
肌细胞生成素
基因克隆
生物信息学
结构特征
系统进化树
Cervus elaphus kansuensis
myogenin
gene cloning
bioinformatics
structural characteristics
phylogenetic evolution
