水牛甲状旁腺素基因克隆与生物信息学分析

发布时间：2018-03-04 12:46

本文选题：水牛　切入点：甲状旁腺素　出处：《中国畜牧兽医》2017年09期 　论文类型：期刊论文

【摘要】：本研究参考黄牛甲状旁腺素(parathyroid hormone,PTH)基因序列设计引物,运用PCR扩增技术对水牛PTH基因进行克隆,并用生物信息学方法对序列进行分析。结果显示,成功克隆了水牛PTH基因完整编码区序列(CDS)348bp,可编码115个氨基酸。该基因编码区序列与黄牛、猪、马、山羊、绵羊和骆驼的同源性分别为99%、93%、91%、97%、96%和92%,表明PTH基因在不同物种间高度保守。系统进化树分析表明,水牛与黄牛PTH基因亲缘关系最近。此外,在水牛PTH基因CDS上共发现了3个碱基突变,其中两个属于同义突变,一个属于错义突变。氨基酸序列分析表明,该蛋白属于碱性、亲水、稳定型蛋白质,其分子式为C_(570)H_(941)N_(167)O_(164)S_8,分子质量为13.01ku。二级结构分析显示,水牛PTH蛋白以α-螺旋和无规则卷曲为主,与三级结构预测结果相一致。亚细胞定位分析显示,水牛PTH蛋白分布在细胞质(26.1%)、线粒体(21.7%)、细胞外(包括细胞壁)(17.4%)、内质网(13.0%)、细胞核(8.7%)、高尔基体(4.3%)和其他部位(8.7%),推测PTH蛋白可能在运输、结合及细胞被膜等方面发挥信号转导和转录调控作用。
[Abstract]:In this study, we designed primers for the sequence of parathyroid hormoney (PTH) gene of bovine parathyroid hormone, cloned the PTH gene of buffalo by PCR amplification technique, and analyzed the sequence by bioinformatics. The complete coding region of buffalo PTH gene was cloned successfully, which can encode 115 amino acids. The sequence of the complete coding region of buffalo PTH gene is similar to that of cattle, pig, horse, goat, yellow cattle, pig, horse and goat. The homology of sheep and camel were 9993and 9197%, respectively, indicating that the PTH gene was highly conserved among different species. Phylogenetic tree analysis showed that the PTH gene of buffalo and yellow cattle had the closest relationship. Three base mutations were found in buffalo PTH gene CDS, two of which were synonymous mutations and one was missense mutation. The amino acid sequence analysis showed that the protein belonged to alkaline, hydrophilic and stable proteins. The molecular formula of this protein is: Che 570H / L / N _ S _ (167O) / S _ (8) and molecular mass 13.01ku.The secondary structure analysis shows that the PTH protein of buffalo is mainly 伪 -helix and irregular curl, which is consistent with the prediction result of tertiary structure. Subcellular localization analysis shows that the molecular weight is 13.01ku.Subcellular localization analysis shows that the protein is mainly 伪 -helix and irregular curl, and is consistent with the predicted results of tertiary structure. The PTH protein of buffalo is distributed in cytoplasm 26.1%, mitochondria 21.7m, extracellular (including cell wall 17.4, endoplasmic reticulum 13.0, nucleus 8.7, Golgi 4.33) and other parts, suggesting that PTH protein may be transported. Binding and cell membrane play a role in signal transduction and transcriptional regulation.
【作者单位】：中国农业科学院广西水牛研究所农业部(广西)水牛遗传繁育重点实验室;
【基金】：国家自然科学基金项目(31660649) 广西科技重大专项(桂科AA16450002) 广西科技公关国合项目(桂科合1504001-3) 水牛基160207 水牛基1705005
【分类号】：Q811.4;S823.83

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