藏绵羊FAM134B基因克隆与表达分析

发布时间：2018-01-21 13:40

  本文关键词： 藏绵羊 FAMB基因 CDS区克隆 生物信息学分析 表达　出处：《浙江农业学报》2017年09期 　论文类型：期刊论文

【摘要】：采用RT-PCR技术克隆了藏绵羊FAM134B(family with sequence similarity 134,member B)基因,利用生物学软件进行相关生物信息学分析,同时用实时荧光定量PCR(real-time fluorescence quantitative PCR,qRT-PCR)研究组织表达水平。克隆获得FAM134B基因全长1 144 bp(Gen Bank登录号:KX580309),开放阅读框1 071 bp,编码356个氨基酸。系统进化树表明,藏绵羊FAM134B和绵羊、山羊关系较近。藏绵羊FAM134B基因编码蛋白的分子式为C_(1717)H_(2696)N_(448)O_(572)S_(12),分子量为39 151.72 u,理论等电点(pI)为4.49,消光系数为24 450,不稳定系数为43,疏水指数为80.31,平均亲水性为-0.422,属不稳定可溶性酸性蛋白质。二级结构主要是α-螺旋(45.51%)、无规卷曲(41.85%)和延伸链(12.64%),属于混合类蛋白质。亚细胞定位和功能分析结果显示,FAM134B编码的蛋白在内质网、细胞质膜概率分别为30.4%和21.7%,主要在蛋白质运输和结合(77.3%)方面起作用。系统发育树表明,藏绵羊FAM134B氨基酸序列与绵羊、山羊同源性非常高。qRT-PCR结果表明,FAM134B基因网胃中表达最高,十二指肠表达量最低。研究结果可为进一步研究藏绵羊FAM134B基因结构和生理功能及其遗传特性提供理论依据。
[Abstract]:Tibetan sheep FAM134B(family with sequence similarity 134 was cloned by RT-PCR technique. Member B) gene was analyzed by bioinformatics using biological software. At the same time, real-time fluorescence quantitative PCR(real-time fluorescence quantitative PCR was used. The total length of FAM134B gene was 1 144 bp(Gen Bank accession number: KX580309). Open reading box 1071 BP, encoding 356 amino acids. Phylogenetic tree showed that Tibetan sheep FAM134B and sheep. The molecular formula of the protein encoded by the FAM134B gene of Tibetan sheep is C _ S _ 1717 / HX _ T _ 2696N _ T _ 4 _ 48 _ O _ S _ S _ S _ 572 _ S _ S _ _ _. The molecular weight is 39 151.72 u, the theoretical isoelectric point Pi is 4.49, the extinction coefficient is 24 450, the unstable coefficient is 43, and the hydrophobic index is 80.31. The average hydrophilicity is -0.422, which belongs to unstable soluble acid protein. The secondary structure is mainly 伪 -helix 45.51). The results of subcellular localization and functional analysis showed that the protein encoded by FAM134B was in the endoplasmic reticulum (ER). The cytosolic membrane probabilities were 30.4% and 21.7, respectively, which mainly played a role in protein transport and binding. The amino acid sequence of FAM134B in Tibetan sheep was highly homologous with sheep and goat. The results of qRT-PCR showed that the expression of FAM134B gene was the highest in reticulated stomach of Tibetan sheep. The results can provide theoretical basis for further study on the structure, physiological function and genetic characteristics of FAM134B gene in Tibetan sheep.
【作者单位】： 甘肃农业大学生命科学技术学院;甘肃省草食动物生物技术重点实验室;甘肃农业大学动物医学院;
【基金】：农业部畜禽遗传资源与种质创新重点实验室开放课题资助(nzdsys2015-1)
【分类号】：S826
【正文快照】： FAM134B(family with sequence similarity134,member B)是最近发现的一种功能基因,其具体的功能还不是太清楚。Kurth等[1]研究发现FAM134B基因调控自主神经元以及疼痛感知神经元。此外,FAM134B基因在人食道鳞状细胞癌变机理方面有着非常重要的意义[2]。高尔基体在蛋白质加工，

本文编号：1451632