水牛溶血磷脂酸受体3基因的克隆及生物信息学分析

doi:10.16431/j.cnki.1671-7236.2018.05.001

摘要/Abstract

摘要：

本试验旨在进行水牛溶血磷脂酸受体3（lysophosphatidic acid receptor 3，LPAR3）基因的克隆及生物信息学分析。以水牛卵巢组织DNA为模板，参考GenBank中公布的水牛LPAR3基因（登录号：XM_006047539.1）序列设计引物，利用PCR扩增水牛LPAR3基因序列，并对其进行生物信息学分析。结果显示，水牛LPAR3基因全序列为1 552 bp，包含1个1 062 bp的CDS序列，编码353个氨基酸。同源性对比结果表明，水牛LPAR3基因编码的氨基酸序列与美洲野牛、黄牛、绵羊、野猪、马、果蝠、白眉猴同源性分别为99.4%、98.9%、98.0%、88.6%、90.0%、90.3%和91.2%，表明LPAR3基因在不同物种间具有较高的保守性。LPAR3蛋白分子式为C₁₈₅₃H₂₈₇₈N₄₇₆O₄₈₆S₃₁，分子质量为40.59 ku，理论等电点（pI）为9.52，不稳定系数为47.05，平均亲水性为0.324，属于碱性不稳定疏水蛋白质；该蛋白质存在7个跨膜结构且无信号肽，属于非分泌蛋白；二级结构分析表明LPAR3以α-螺旋、无规则卷曲和延伸链为主，其中α-螺旋占48.16%，无规则卷曲占41.36%，延伸链占10.48%，属于全α类蛋白质，与三级结构预测结果一致；亚细胞定位分析发现，LPAR3蛋白分布在等离子体膜（56.5%）、内质网（26.1%）、空泡（8.7%）、高尔基体（4.3%）和细胞核（4.3%）中，推测可能在运输和结合及嘌呤和嘧啶等方面发挥转运蛋白和信号转导等作用。本试验结果可为今后深入探讨LPAR3基因功能奠定基础。

关键词: 水牛; 溶血磷脂酸受体3; 基因克隆; 生物信息学

Abstract:

The study was aimed to clone and analyze the lysophosphatidic acid receptor 3 (LPAR3) gene of buffalo with bioinformatics method.The ovary was collected from buffalo,DNA samples was extracted and the primers were designed according to the sequence of the buffalo LPAR3 gene (GenBank accession No.:XM_006047539.1).And LPAR3 gene was cloned and analyzed by PCR amplification and bioinformatics softwares.The results showed that LPAR3 gene was 1 552 bp in length containing a CDS sequence of 1 062 bp that encoded 353 amino acids.Sequence homology analysis indicated that the buffalo LPAR3 showed 99.4%,98.9%,98.0%,88.6%,90.0%,90.3% and 91.2% identity with that of Bison bison,Bos taurus,Ovis aries,Sus scrofa,Equus caballus,Rousettus and Cercocebus atys,which was consistent with phylogenetic tree analysis.The formula of the LPAR3 protein was C₁₈₅₃H₂₈₇₈N₄₇₆O₄₈₆S₃₁,and the molecular weight,theory isoelectric point,instability index and grand average of hydrophobicity was 40.59 ku,9.52,47.05 and 0.324,respectively,indicating that it was an alkaline unstable hydrophobic protein.There were 7 transmembrane domain but no signal peptide which showed it was a non-secretory protein.The secondary structure analysis showed that the percentage of α-helices,random coil and extended strand was 48.16%,41.36% and 10.48%,respectively.Protein subcellular localization analysis found that LPAR3 distributed in the plasma membrane (56.5%),endoplasmic reticulum (26.1%),vacuole (8.7%),Golgi apparatus (4.3%) and the nucleus (4.3%), and might play a role in the transport and combination,purines and pyrimidines of transporters and signal transduction.Thus,those results would lay a foundation for clarifying the mechanism of the gene performance in buffaloes.

Key words: buffalo; lysophosphatidic acid receptor 3 (LPAR3); gene cloning; bioinformatics

中图分类号:

庞春英, 梁莎莎, 马小娅, 邓廷贤, 陆杏蓉, 段安琴, 黄韵琪, 梁贤威. 水牛溶血磷脂酸受体3基因的克隆及生物信息学分析[J]. 《中国畜牧兽医》, 2018, 45(5): 1127-1136.

PANG Chunying, LIANG Shasha, MA Xiaoya, DENG Tingxian, LU Xingrong, DUAN Anqin, HUANG Yunqi, LIANG Xianwei. Cloning and Bioinformatics Analysis of LPAR3 Gene in Buffalo[J]. , 2018, 45(5): 1127-1136.

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