新疆哈萨克绵羊MHC Ⅰ类分子基因的克隆与生物信息学分析

发布时间：2018-03-07 02:14

  本文选题：新疆哈萨克绵羊　切入点：主要组织相容性复合体(MHC)Ⅰ类分子基因　出处：《黑龙江畜牧兽医》2017年17期 　论文类型：期刊论文

【摘要】：为了研究新疆哈萨克绵羊主要组织相容性复合体(MHC)Ⅰ类分子基因序列的多态性,采用PCR和测序的方法分别克隆新疆哈萨克绵羊MHCⅠ类分子基因重链和轻链的c DNA全长,并进行生物信息学分析。结果表明:成功扩增出新疆哈萨克绵羊MHCⅠ类分子基因重链、轻链,大小分别为1 107 bp、357 bp;通过生物信息学软件预测蛋白质分子质量分别为90 ku和29.694 ku;等电点(p I)分别为5.00,5.23;重链富含G、C碱基,有两段跨膜螺旋,最大疏水指数为3.622,最小疏水指数为-2.922,预测整条肽链表现为亲水性,在1~84位碱基之间存在信号肽;轻链四种碱基含量基本一致,也有两段跨膜螺旋,在1~63位碱基之间存在信号肽。新疆哈萨克绵羊MHCⅠ类分子基因编码蛋白质的二级结构以无规则卷曲元件的含量最高,该蛋白质的三级结构由一条α链,另一条为β链通过非共价键组成,由α1、α2组成抗原结合槽;NCBI数据库中不同地区的哈萨克绵羊的MHCⅠ类分子基因的同源性在89.87%~99.91%之间,与数据库中牛的MHCⅠ类分子基因同源性达到了98.88%,轻链MHCⅠ类分子基因还与猪和人的同源性相对较高。
[Abstract]:In order to study the polymorphism of class I gene sequence of Xinjiang Kazak sheep main histocompatibility complex (MHC), the length of c DNA of MHC class I heavy chain and light chain of Xinjiang Kazak sheep was cloned by PCR and sequencing, respectively. The results of bioinformatics analysis showed that the heavy chain and light chain of MHC class I gene in Xinjiang Kazakh sheep were successfully amplified. The molecular weight of the protein was estimated by bioinformatics software to be 90 ku and 29.6ku94, the isoelectric point I was 5.00 ~ 5.23, respectively. The heavy chain was rich in G ~ (+) C base and had two transmembrane helices. The maximum hydrophobic index was 3.622, and the minimum hydrophobic index was -2.922. It was predicted that the whole peptide chain was hydrophilic, and there were signal peptides between the 1 ~ 84 bases, and the four bases in the light chain were basically the same, and there were also two transmembrane helices. In Xinjiang Kazak sheep, the secondary structure of the protein encoded by MHC class I gene was the highest in the irregular curl element, and the tertiary structure of the protein consisted of a 伪 chain. The other is that the 尾 chain is composed by non-covalent bonds, and the homology of MHC class I gene of Kazakh sheep in different regions in the database of 伪 1, 伪 2 antigen-binding trough is between 89.87% and 99.91%. The homology of bovine MHC class I gene was 98.88, and that of light chain MHC class I gene was higher than that of pig and human.
【作者单位】： 石河子大学动物科技学院;
【基金】：国家自然科学基金项目(31172303)
【分类号】：S826
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本文编号：1577492