北极狐及乌苏里貉抑制素α亚基基因克隆及生物信息学分析
发布时间：2018-11-21 13:22
【摘要】：试验旨在克隆北极狐及乌苏里貉抑制素α(inhibinα,INHα)亚基基因并对其进行生物信息学分析。根据GenBank中犬科INHα预测mRNA序列(登录号:XM_545660.5)设计1对引物,用RT-PCR技术从北极狐及乌苏里貉的卵巢组织中扩增出INHα亚基基因,同时将其插入到克隆载体中,进行测序及生物信息学分析。测序结果表明,北极狐及乌苏里貉的INHα亚基基因CDS序列全长为1 107bp,编码369个氨基酸。北极狐及乌苏里貉的INHα亚基基因与犬的同源性最高,分别为97.9%与97.6%。系统进化树分析表明,北极狐及乌苏里貉与犬亲缘关系较近,同时也说明INHα亚基基因在不同物种及进化过程中具有高度保守性。对INHα亚基蛋白的高级结构预测发现,由于半胱氨酸间形成的二硫键导致其采用"蝴蝶形"或"开放手"构型,其中α-螺旋形成分子的"手腕"结构,β-折叠形成分子的"手指"结构。本研究成功克隆了北极狐及乌苏里貉的INHα亚基基因,同时进行了系统的生物信息学分析,为今后研究抑制素在卵母细胞-颗粒细胞同步发育过程中的生物学功能奠定了基础。
[Abstract]:The aim of this study was to clone inhibin 伪 (INH 伪) subunit genes from arctic fox and raccoon dog and to analyze them by bioinformatics. According to the INH 伪 predicted mRNA sequence (accession number: XM_545660.5) in GenBank, a pair of primers were designed. The INH 伪 subunit gene was amplified from the ovarian tissues of Arctic fox and raccoon dog by RT-PCR technique and inserted into the clone vector. Sequencing and bioinformatics analysis were performed. The results of sequencing showed that the CDS sequence of INH 伪 subunit gene of arctic fox and raccoon was 1 107 BP, encoding 369 amino acids. The homology of INH 伪 subunit gene between arctic fox and raccoon dog was 97.9% and 97.6%, respectively. Phylogenetic tree analysis showed that arctic fox and raccoon dog were closely related to dogs, and that INH 伪 subunit gene was highly conserved in different species and evolution process. The advanced structure prediction of INH 伪 subunit protein showed that the disulfide bond formed between cysteine resulted in the "butterfly" or "open hand" configuration, in which 伪 -helix formed the "wrist" structure of the molecule. 尾-fold to form the "finger" structure of the molecule. In this study, the INH 伪 subunit genes of arctic fox and raccoon dog were cloned successfully, and the bioinformatics analysis was carried out systematically, which laid a foundation for studying the biological function of inhibin in the process of oocyte granulosa cell development in the future.
【作者单位】： 中国农业科学院特产研究所;中国农业科学院特种经济动物分子生物学重点实验室;
【基金】：中国农业科学院创新工程(特种经济动物繁殖创新团队);中国农业科学院基本科研业务费专项(1610342017025) 吉林省国际合作项目(20170414049GH) 国家自然基金面上项目(31772606)
【分类号】：S865.2
本文编号：2347080
[Abstract]:The aim of this study was to clone inhibin 伪 (INH 伪) subunit genes from arctic fox and raccoon dog and to analyze them by bioinformatics. According to the INH 伪 predicted mRNA sequence (accession number: XM_545660.5) in GenBank, a pair of primers were designed. The INH 伪 subunit gene was amplified from the ovarian tissues of Arctic fox and raccoon dog by RT-PCR technique and inserted into the clone vector. Sequencing and bioinformatics analysis were performed. The results of sequencing showed that the CDS sequence of INH 伪 subunit gene of arctic fox and raccoon was 1 107 BP, encoding 369 amino acids. The homology of INH 伪 subunit gene between arctic fox and raccoon dog was 97.9% and 97.6%, respectively. Phylogenetic tree analysis showed that arctic fox and raccoon dog were closely related to dogs, and that INH 伪 subunit gene was highly conserved in different species and evolution process. The advanced structure prediction of INH 伪 subunit protein showed that the disulfide bond formed between cysteine resulted in the "butterfly" or "open hand" configuration, in which 伪 -helix formed the "wrist" structure of the molecule. 尾-fold to form the "finger" structure of the molecule. In this study, the INH 伪 subunit genes of arctic fox and raccoon dog were cloned successfully, and the bioinformatics analysis was carried out systematically, which laid a foundation for studying the biological function of inhibin in the process of oocyte granulosa cell development in the future.
【作者单位】： 中国农业科学院特产研究所;中国农业科学院特种经济动物分子生物学重点实验室;
【基金】：中国农业科学院创新工程(特种经济动物繁殖创新团队);中国农业科学院基本科研业务费专项(1610342017025) 吉林省国际合作项目(20170414049GH) 国家自然基金面上项目(31772606)
【分类号】：S865.2
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