水牛LPIN1基因克隆及序列分析

doi:10.16431/j.cnki.1671-7236.2018.10.001

摘要/Abstract

摘要：

试验旨在克隆获得水牛脂素1（LPIN1）基因，并对其进行生物信息学分析，为揭示该基因在水牛脂肪沉积、生殖发育和泌乳调控中的作用奠定基础。本研究以水牛卵巢组织cDNA为模板，PCR扩增获得了LPIN1基因CDS区全长后测序，并结合生物信息学分析方法预测及分析蛋白质理化性质、二级结构及三级结构等。结果表明，水牛LPIN1基因编码区长2 793 bp，编码930个氨基酸。MegAlign软件分析显示，水牛LPIN1基因核苷酸序列与水牛（预测）、牦牛、黄牛、山羊、藏羚羊、绵羊、猪、骆驼、人和小鼠LPIN1基因的同源性分别为99.6%、97.9%、97.7%、97.5%、97.4%、97.1%、89.9%、89.8%、86.2%和83.5%；水牛lipin1蛋白氨基酸序列与黄牛、牦牛、山羊、藏羚羊、骆驼、猪及人的同源性分别为99%、99%、99%、99%、94%、94%及90%。应用Mega 5.0软件构建系统进化树发现，水牛与黄牛的亲缘关系最近，其次为绵羊和山羊，LPIN1基因在不同物种及进化的过程中具有高度保守性。对lipin1蛋白分析发现，其二级结构由α-螺旋、β-折叠、T-转角和无规则卷曲组成；蛋白呈弱酸性，无信号肽，亚细胞主要定位于细胞核中，存在Lipin_N、LNS2和AF1Q等结构域，其中Lipin_N、LNS2为保守结构域。

关键词: 水牛; 脂素1基因(LPIN1); 克隆; 生物信息学分析

Abstract:

In order to clarify the effect of LPIN1 gene on the fat deposition,reproductive performance and lactation regulation of buffalo,LPIN1 gene was cloned and analyzed by bioinformatics.cDNA was extracted from buffalo ovary,and then LPIN1 gene CDS region was cloned and sequenced.The physical and chemical properties,secondary structure and tertiary structure of lipin1 protein were predicted and analyzed by bioinformatics software.The results showed that the coding region of buffalo LPIN1 gene was 2 793 bp,which encoded 930 amino acids.The buffalo LPIN1 gene nucleotide sequence shared 99.6%,97.9%,97.7%,97.5%,97.4%,97.1%,89.9%,89.8%,86.2% and 83.5% of homology with that of Bubalus bubalis(predicted),Bos mutus,Bos taurus,Capra hircus,Pantholops hodgsonii,Ovis aries,Sus scrofa,Camelus bactrianus,Homo sapiens and Mus musculus,respectively.The buffalo lipin1 protein shared 99%,99%,99%,99%,94%,94% and 90% of similar amino acids sequence with that of Bos taurus,Bos mutus,Capra hircus,Pantholops hodgsonii,Camelus bactrianus,Sus scrofa and Homo sapiens.The results of phylogenetic tree constructed by Mega 5.0 showed that LPIN1 gene in buffalo had a closest relation with Bos taurus,followed by Ovis aries and Capra hircus,indicating LPIN1 gene was highly conserved in different species.The secondary structure of the lipin1 protein was formed with α-helix,β-turn,T-turn and random coil.lipin1 protein was weakly acidic,which mianly located in the nucleus,and had Lipin_N,LNS2 and AF1Q domain and no signal peptide.The Lipin_N and LNS2 were the conservation domain.

Key words: buffalo; LPIN1 gene; cloning; bioinformatics analysis

中图分类号:

Q785

段安琴, 庞春英, 邓廷贤, 陆杏蓉, 马小娅, 梁莎莎, 梁贤威. 水牛LPIN1基因克隆及序列分析[J]. 《中国畜牧兽医》, 2018, 45(10): 2653-2660.

DUAN Anqin, PANG Chunying, DENG Tingxian, LU Xingrong, MA Xiaoya, LIANG Shasha, LIANG Xianwei. Cloning and Sequence Analysis of Buffalo LPIN1 Gene[J]. , 2018, 45(10): 2653-2660.

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